Salts of compounds and their crystal forms

Abstract

The present invention belongs to the field of pharmacy, and provides a pharmaceutically acceptable salt of compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine, a crystal form thereof, a solvate and a crystal form thereof, a pharmaceutical composition thereof, and a preparation method and application thereof.

Description

[0001] This application is a divisional application of Chinese patent application No. 201880052524.1, with an application date of August 22, 2018 and a priority date of August 22, 2017. Technical Field

[0002] The present invention belongs to the field of pharmacy, and provides a pharmaceutically acceptable salt of the compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine, a crystal form thereof, a solvate and a crystal form thereof, a pharmaceutical composition thereof, and a preparation method and application thereof. Background Art

[0003] Spleen tyrosine kinase (Syk) is a non-receptor tyrosine kinase that plays an important role in immune receptor- and integrin-mediated signaling in many cells, including B cells, macrophages, monocytes, mast cells, eosinophils, basophils, neutrophils, dendritic cells, T cells, natural killer cells, platelets, and osteoclasts.

[0004] Syk is necessary for B cell activation mediated by B cell receptor signaling. Syk is activated after binding to phosphorylated B cell receptors, which in turn initiates the B cell receptor signaling pathway, thereby mediating a variety of biological effects. This signaling pathway in turn depends on the developmental stage of B cells. The intensity and duration of B cell receptor signals must be strictly regulated. Abnormal B cell receptor signaling pathways can lead to dysregulated B cell activation or the formation of pathological autoantibodies, resulting in multiple autoimmune or inflammatory diseases. Mice deficient in Syk have impaired B cell maturation, reduced immunoglobulin production, weakened immune responses independent of T cells, and significantly weakened calcium ion signals caused by B cell receptor activation.

[0005] A large amount of evidence shows that B cells and the humoral immune system play an important role in the pathogenesis of autoimmune and inflammatory diseases. Autoantibodies and the immune complexes they produce are involved in the pathological processes of autoimmune and inflammatory diseases. The pathological response to these antibodies depends on the activation of the Syk-mediated signaling pathway through the Fc receptor. Due to the role of Syk in B cell activation and its role in the Fc signaling pathway, B cell-mediated pathological activities, including the production of autoantibodies, can be inhibited by inhibiting Syk. Therefore, the production of autoantibodies can be inhibited by inhibiting the activity of the intracellular Syk enzyme, thereby achieving the purpose of treating autoimmune diseases.

[0006] Therefore, inhibiting the activity of Syk can be used to treat autoimmune diseases and inflammatory diseases such as systemic lupus erythematosus, rheumatoid arthritis, Sjögren's syndrome, IgA nephropathy, autoimmune hemolytic anemia, multiple sclerosis, idiopathic thrombocytopenic purpura, myasthenia gravis, allergic rhinitis, chronic obstructive pulmonary disease, adult respiratory distress syndrome, and asthma. Moreover, it is reported that Syk mediates the ligand-independent signaling pathway mediated by the B cell receptor, which is also an important signaling pathway for the survival of known B cells. Therefore, inhibiting the activity of Syk can be used to treat cancer, preferably hematological malignancies such as lymphoma, leukemia and multiple myeloma.

[0007] The related compound of the present invention (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine has the effect of effectively inhibiting the activity of Syk kinase. Therefore, it can be used for the treatment of diseases that respond to the inhibition of Syk kinase activity, such as the treatment of autoimmune diseases, inflammatory diseases and cancer.

[0008] The phenomenon that a compound can exist in two or more crystal structures is called polymorphism. Many compounds can exist in multiple crystal forms or in amorphous solid forms. Before the discovery of polymorphism of a compound, it is difficult to predict (1) whether a particular compound has polymorphism; (2) how to produce these unknown polymorphs; and (3) what the properties of these polymorphs will be, such as stability. See J. Bernstein "Polymorphism in Molecular Crystals", Oxford University Press, (2002).

[0009] Salt formation will not change the biological activity of the compound itself, but it can change the physical and chemical properties of the compound, such as solubility, stability, crystallinity, polymorphism, etc. However, it is impossible to predict which specific properties will change and the extent of the change. It is also impossible to predict whether a certain compound or its salt formed with an acid / base can be crystalline or amorphous, what kind of crystal will be formed, how a certain crystal is formed, and whether the formed crystal has a certain property.

[0010] Since the properties of solids depend on the structure and the properties of the compound itself, different solid forms of compounds often exhibit different physical and chemical properties and different biopharmaceutical properties. Differences in chemical properties can be determined, analyzed and compared by a variety of analytical techniques, and these differences can ultimately be used to distinguish between the different solid forms that exist. Differences in physical properties such as solubility and biopharmaceutical properties such as bioavailability are also important in describing the solid forms of pharmaceutical compounds. Similarly, in the development of pharmaceutical compounds, such as the compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine, salts, new crystal forms and amorphous forms of pharmaceutical compounds are also important.

[0011] Patent application WO2012167733A1 discloses the compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine and its preparation method. Summary of the invention

[0012] Overview

[0013] After extensive exploration and research, we found that the compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine can be prepared into various pharmaceutically acceptable salts, and its chemical structure is shown in Formula A. The results show that the solubility of the salt of Formula A is significantly increased relative to its free base, which is beneficial to improving the pharmacokinetic characteristics of the compound and increasing the bioavailability of the compound in vivo. We also found that the salt of Formula A can exist in different crystalline forms (i.e., polymorphs) and can form solvates with certain solvents. We have conducted extensive research on the polymorphs of the salt of Formula A and determined and prepared crystalline forms that meet pharmaceutical needs. Based on these studies, the present invention provides multiple crystalline forms of pharmaceutically acceptable salts of compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine, their solvates and crystalline forms thereof, which are respectively designated as Form AI, Form BI, Form B-II, Form B-III, Form CI, Form C-II and Form C-III.

[0014]

[0015] Where n is 0.5 or 1;

[0016] M is a pharmaceutically acceptable acid molecule.

[0017] In one aspect, the present invention provides a polymorph of the salt of formula A or a solvate thereof, which has the characteristics of good crystallinity, high solubility and good stability.

[0018] First, the present invention provides a pharmaceutically acceptable salt of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine.

[0019] Secondly, the present invention provides a pharmaceutically acceptable salt of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine as shown in formula A, that is, the salt of formula A.

[0020] Secondly, the present invention provides pharmaceutically acceptable salts of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine as shown in formula A, which are acetate, p-toluenesulfonate and malate, respectively.

[0021] Secondly, the present invention provides pharmaceutically acceptable salts of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine as shown in formula A, which are monoacetate, p-toluenesulfonate, monomalate and hemimalate, respectively.

[0022] Secondly, the present invention provides a crystal of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine monoacetate, i.e., a crystal of formula A salt (wherein n is 1 and M is acetic acid) in form AI.

[0023] Secondly, the present invention provides a BI type crystal of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine p-toluenesulfonate, that is, a crystalline form BI of the salt of formula A (wherein n is 1 and M is p-toluenesulfonic acid).

[0024] Secondly, the present invention provides a B-II type crystal of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine p-toluenesulfonate, that is, a crystal form B-II of the salt of formula A (wherein n is 1 and M is p-toluenesulfonic acid).

[0025] Secondly, the present invention provides a solvate of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine p-toluenesulfonate, which is a hydrate of the salt of formula A (wherein n is 1 and M is p-toluenesulfonic acid).

[0026] Secondly, the present invention provides a hydrate of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine p-toluenesulfonate, which is a hemihydrate of the salt of formula A (wherein n is 1 and M is p-toluenesulfonic acid).

[0027] Secondly, the present invention provides a hydrate of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine p-toluenesulfonate, which is a hemihydrate of the salt of formula A (wherein n is 1 and M is p-toluenesulfonic acid), which is a crystalline form B-III.

[0028] Secondly, the present invention provides a CI type crystal of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine monomalate, that is, a crystal form CI of the salt of formula A (wherein n is 1 and M is malic acid).

[0029] Secondly, the present invention provides a solvate of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine malate, which is a hydrate of the salt of formula A (wherein n is 0.5 and M is malic acid).

[0030] Secondly, the present invention provides hydrates of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine malate, which are respectively a hydrate and a monohydrate of the salt of formula A (wherein n is 0.5 and M is malic acid) containing 0.75 molecules of water.

[0031] Secondly, the present invention provides a hydrate of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine malate, which is a hydrate of the salt of formula A (wherein n is 0.5 and M is malic acid) containing 0.75 molecules of water, and is a crystalline form C-II.

[0032] Secondly, the present invention provides a hydrate of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine malate, which is a monohydrate of the salt of formula A (wherein n is 0.5 and M is malic acid), which is a crystalline form C-III.

[0033] On the other hand, the present invention provides methods for preparing a salt of formula A or a solvate thereof or a crystalline form of a salt of formula A or a solvate thereof (e.g., form AI, form BI, form B-II, form B-III, form CI, form C-II and form C-III), which preparation methods are repeatable and easy to operate.

[0034] In another aspect, the present invention provides a pharmaceutical composition, which contains an effective amount of a salt of formula A or a solvate thereof, or one or more crystalline forms of a salt of formula A or a solvate thereof (e.g., form AI, form BI, form B-II, form B-III, form CI, form C-II, and form C-III), and the remainder is at least one pharmaceutically acceptable carrier.

[0035] The present invention also provides a method for treating a disease that responds to inhibition of Syk kinase activity. The method comprises administering to an individual in need thereof an effective amount of one or more salts of Formula A of the present invention or a solvate thereof or a crystalline form of a salt of Formula A or a solvate thereof, such as Form AI, Form BI, Form B-II, Form B-III, Form CI, Form C-II or Form C-III.

[0036] The present invention also provides the use of a salt of formula A or a solvate thereof or a crystalline form of a salt of formula A or a solvate thereof (such as crystalline form AI, crystalline form BI, crystalline form B-II, crystalline form B-III, crystalline form CI, crystalline form C-II or crystalline form C-III) for preparing a drug, wherein the drug is used to treat diseases responsive to inhibition of Syk kinase activity, such as autoimmune diseases, inflammatory diseases and cancer (preferably hematological malignancies), such as systemic lupus erythematosus, rheumatoid arthritis, Sjögren's syndrome, IgA nephropathy, autoimmune hemolytic anemia, idiopathic thrombocytopenic purpura, multiple sclerosis, myasthenia gravis, allergic rhinitis, chronic obstructive pulmonary disease, adult respiratory distress syndrome, asthma, lymphoma (such as B-cell lymphoma, T-cell lymphoma), leukemia (such as chronic lymphocytic leukemia, acute lymphocytic leukemia and acute myeloid leukemia) and multiple myeloma. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 The powder X-ray diffraction pattern of the crystalline form AI of the salt of formula A is shown, with the horizontal axis (X-axis) being the diffraction angle 2θ and the vertical axis (Y-axis) being the diffraction intensity.

[0038] Figure 2 The differential scanning calorimetry graph of Form AI of the salt of Formula A is shown, with the horizontal axis (X-axis) being temperature and the vertical axis (Y-axis) being heat flow.

[0039] Figure 3 Thermogravimetric analysis graph showing Form AI of the salt of Formula A, wherein the horizontal axis (X-axis) is temperature and the vertical axis (Y-axis) is weight percentage.

[0040] Figure 4 The powder X-ray diffraction pattern of the crystalline form BI of the salt of formula A is shown, with the horizontal axis (X-axis) being the diffraction angle 2θ and the vertical axis (Y-axis) being the diffraction intensity.

[0041] Figure 5 A differential scanning calorimetry graph of Form BI of the salt of Formula A is shown, with the horizontal axis (X-axis) being temperature and the vertical axis (Y-axis) being heat flow.

[0042] Figure 6 A thermogravimetric analysis graph showing Form BI of the salt of Formula A, wherein the horizontal axis (X-axis) is temperature and the vertical axis (Y-axis) is weight percentage.

[0043] Figure 7 The powder X-ray diffraction pattern of the crystalline form B-II of the salt of formula A is shown, wherein the horizontal axis (X-axis) is the diffraction angle 2θ and the vertical axis (Y-axis) is the diffraction intensity.

[0044] Figure 8 The differential scanning calorimetry diagram and thermogravimetric analysis diagram of Form B-II of the salt of Formula A are shown, with the horizontal axis (X-axis) being temperature and the vertical axis (Y-axis) being heat flow and weight percentage.

[0045] Fig. 9 The powder X-ray diffraction pattern of the crystalline form B-III of the hemihydrate of the salt of formula A is shown, wherein the horizontal axis (X-axis) is the diffraction angle 2θ and the vertical axis (Y-axis) is the diffraction intensity.

[0046] Fig.10 The differential scanning calorimetry diagram and thermogravimetric analysis diagram of the hemihydrate form B-III of the salt of formula A are shown, with the horizontal axis (X-axis) being temperature and the vertical axis (Y-axis) being heat flow and weight percentage.

[0047] Fig.11 The powder X-ray diffraction pattern of the crystalline form CI of the salt of formula A is shown, with the horizontal axis (X-axis) being the diffraction angle 2θ and the vertical axis (Y-axis) being the diffraction intensity.

[0048] Fig.12 A differential scanning calorimetry graph of Form CI of the salt of Formula A is shown, with the horizontal axis (X-axis) being temperature and the vertical axis (Y-axis) being heat flow.

[0049] Fig.13Thermogravimetric analysis graph showing Form CI of the salt of Formula A, with the horizontal axis (X-axis) being temperature and the vertical axis (Y-axis) being weight percentage.

[0050] Fig.14 The powder X-ray diffraction pattern of the crystalline form C-II of the hydrate of the salt of formula A is shown, wherein the horizontal axis (X-axis) is the diffraction angle 2θ and the vertical axis (Y-axis) is the diffraction intensity.

[0051] Fig.15 A differential scanning calorimetry graph showing Form C-II of the hydrate of the salt of Formula A, with the horizontal axis (X-axis) being temperature and the vertical axis (Y-axis) being heat flow.

[0052] Fig.16 Thermogravimetric analysis chart of Form C-II of the hydrate of the salt of Formula A, wherein the horizontal axis (X-axis) is temperature and the vertical axis (Y-axis) is weight percentage.

[0053] Fig.17 The powder X-ray diffraction pattern of the crystalline form C-III of the monohydrate of the salt of formula A is shown, wherein the horizontal axis (X-axis) is the diffraction angle 2θ and the vertical axis (Y-axis) is the diffraction intensity.

[0054] Fig.18 A differential scanning calorimetry graph showing Form C-III of the monohydrate of the salt of Formula A, with the horizontal axis (X-axis) being temperature and the vertical axis (Y-axis) being heat flow.

[0055] Fig.19 Thermogravimetric analysis chart showing the crystalline form C-III of the monohydrate of the salt of Formula A, wherein the horizontal axis (X-axis) is temperature and the vertical axis (Y-axis) is weight percentage. DETAILED DESCRIPTION

[0056] definition

[0057] Unless otherwise stated, the following abbreviations or terms used in this application (including the specification and claims) have the definitions given below. It must be noted that the singular forms used in this specification and the appended claims also include the plural forms, unless the context clearly indicates otherwise.

[0058] As used herein, the "crystalline form of the present invention" refers to the crystalline form AI, crystalline form BI, crystalline form B-II, crystalline form B-III, crystalline form CI, crystalline form C-II or crystalline form C-III of the salt of Formula A or its solvate, or a mixture of several of them. "Crystal form", "crystalline form" and "polymorph" are used interchangeably herein.

[0059] As used herein, "Salt of Formula A" refers to a salt having the following chemical structure of Formula A (also referred to as Salt A):

[0060]

[0061] Where n is 0.5 or 1;

[0062] M is a pharmaceutically acceptable acid molecule.

[0063] The “C 1-6 "Alkyl alcohol" refers to a fully saturated straight or branched chain alkyl alcohol having 1, 2, 3, 4, 5 or 6 carbon atoms. Examples include, but are not limited to, methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, tert-butanol, n-pentanol, isopentanol, n-hexanol, etc.

[0064] As used herein, "halogenated alkanes having less than three carbon atoms" refer to fully saturated hydrocarbons having 1 or 2 carbon atoms, which are substituted with one or more halogen atoms selected from F, Cl, Br or I. Examples thereof include dichloromethane, chloroform, carbon tetrachloride, 1,2-dichloroethane and the like.

[0065] As used herein, "about" refers to a deviation of up to ±10% from the specifically given numerical value.

[0066] As used herein, "substantially free of other crystalline forms" means that the content of the other crystalline forms is less than 50%, preferably less than 40%, preferably less than 30%, preferably less than 20%, preferably less than 10%, preferably less than 5%, and preferably less than 1% by weight of the total weight of the crystalline forms.

[0067] As used herein, "solution" refers to a mixture of one or more solutes in one or more solvents for a certain purpose. Solution is intended to include homogeneous mixtures, and multiphase mixtures, such as slurries or other suspended mixtures containing insoluble matter.

[0068] As used herein, "organic solvent" refers broadly to any suitable organic solvent for the purpose herein.

[0069] As used herein, "dissolving solvent" refers to any appropriate organic solvent that can partially or completely dissolve the solute under appropriate conditions, such as appropriate amount, appropriate temperature, such as room temperature or elevated temperature.

[0070] As used herein, "anti-solvent" refers to any suitable organic solvent in which the solubility of a substance is less than that of the solvent.

[0071] As used herein, "pharmaceutically acceptable salts" include, but are not limited to, salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, phosphorous acid, sulfuric acid, sulfurous acid, nitric acid, and the like, and salts formed with organic acids such as malate, maleate, mandelate, fumarate, tartrate, succinate, citrate, aspartate, glutamate, 2-hydroxy-2-phenylpropionate, gluconate, lactate, camphorsulfonate, methanesulfonate, ethanesulfonate, naphthylate, p-toluenesulfonate, 2-hydroxyethanesulfonate, β-hydroxybutyrate, benzoate, salicylate, and alkyl carboxylates such as acetate, propionate, stearate, HOOC-(CH2)n-COOH (wherein n is 0-4), and the like.

[0072] The "effective amount" of the salt of formula A and the crystalline form, solvate and crystalline form thereof refers to the amount administered to an individual that can effectively alleviate or improve diseases responsive to the inhibition of Syk kinase activity, such as autoimmune diseases, inflammatory diseases and cancers (preferably hematological malignancies). The subject of administration can be a human or an animal subject, wherein the diseases responsive to the inhibition of Syk kinase activity, such as autoimmune diseases, inflammatory diseases and cancers (preferably hematological malignancies) can be, but are not limited to, systemic lupus erythematosus, rheumatoid arthritis, Sjögren's syndrome, IgA nephropathy, autoimmune hemolytic anemia, idiopathic thrombocytopenic purpura, multiple sclerosis, myasthenia gravis, allergic rhinitis, chronic obstructive pulmonary disease, adult respiratory distress syndrome, asthma, lymphoma (such as B-cell lymphoma, T-cell lymphoma), leukemia (such as chronic lymphocytic leukemia, acute lymphocytic leukemia and acute myeloid leukemia) and multiple myeloma, etc. The "effective amount" will vary depending on many factors, such as the compound, the disease state being treated, the severity of the disease being treated, the age and relevant health of the individual, the route and form of administration, and the judgment of the attending physician or veterinary practitioner.

[0073] As used herein, "individual" means mammals and non-mammals. Mammals mean any member of the class mammals, including but not limited to humans; non-human primates such as chimpanzees and other apes and monkeys; farm animals such as cattle, horses, sheep, goats and pigs; domestic animals such as rabbits, dogs and cats; laboratory animals, including rodents such as rats, mice and guinea pigs; etc. Examples of non-mammals include but are not limited to birds, etc. The term "individual" does not indicate a particular age or sex. DETAILED DESCRIPTION OF THE INVENTION

[0075] The present invention provides a pharmaceutically acceptable salt of the compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine and a crystalline form thereof, a solvate thereof and a crystalline form thereof.

[0076] The crystal form provided by the present invention has the characteristics of good crystallinity, high solubility and good stability. The crystal form of the present invention has good reproducibility and can realize repeated amplification production; and it is stable in common preparations, so it is convenient for preparation production and use in the treatment of diseases. In addition, the crystal form of the present invention has high purity and less solvent residue, which meets the quality requirements of raw materials, such as the requirements of ICH Q3A.

[0077] Those skilled in the art can verify the above advantages of the crystal form of the present invention based on the test methods disclosed in the pharmacopoeia, its modified methods or conventional methods in the art.

[0078] As described herein, the crystal forms of the present invention can be identified by one or more solid-state analysis methods. For example, the crystal forms of the present invention can be identified by one or more methods, such as powder X-ray diffraction, lattice parameters of single crystals, Fourier transform infrared spectroscopy, differential scanning calorimetry analysis data and / or thermogravimetric curves. And if the identification analysis results of one of the methods are consistent with the crystal forms of the present invention, it does not mean that the identification results of any other method are consistent with the crystal forms of the present invention.

[0079] As described herein, the new crystal form can be identified by powder X-ray diffraction spectrum. However, those skilled in the art know that the peak intensity and / or peak conditions of powder X-ray diffraction may vary due to different experimental conditions, such as different diffraction test conditions and / or preferred orientations, etc. At the same time, due to the different precision of different instruments, the measured 2θ value will have an error of about ±0.2 2θ. However, it is known that the relative intensity value of the peak depends more on certain properties of the measured sample than the position of the peak, such as the size of the crystals in the sample, the orientation of the crystals, and the purity of the analyzed material, so the displayed peak intensity deviation is about ±20% or more. It is possible. However, despite the existence of experimental errors, instrument errors, preferred orientations, etc., those skilled in the art can still obtain sufficient information for identifying crystal form AI and other various crystal forms of the present invention from the XRPD data provided herein.

[0080] Crystal AI

[0081] The present invention provides a crystalline form AI of a salt of formula A (wherein n is 1 and M is acetic acid).

[0082] In some embodiments, the crystalline form AI of the salt of formula A can be identified by X-ray powder diffraction. In some embodiments, the powder X-ray diffraction characteristic diffraction angle (2θ) of the crystalline form AI of the salt of formula A includes 6.2, 9.4, 16.3, 17.2 and 19.0 degrees, and the measured 2θ value has an error of about ± 0.2 2θ.

[0083] In some embodiments, the powder X-ray diffraction characteristic diffraction angles (2θ) of Form AI of Formula A salt include 6.2, 9.4, 10.8, 16.3, 17.2, 19.0, 20.1, 21.7, 24.6 and 29.2 degrees, and the measured 2θ value has an error of about ±0.2 2θ.

[0084] In some embodiments, the powder X-ray diffraction characteristic diffraction angles (2θ) of Form AI include 6.2, 9.4, 10.8, 12.1, 15.8, 16.3, 17.2, 19.0, 20.1, 21.7, 22.7, 24.6, 25.4 and 29.2 degrees, and the measured 2θ values ​​have an error of about ±0.2 2θ.

[0085] In some embodiments, the powder X-ray diffraction characteristic diffraction angle (2θ) of Form AI of the Formula A salt includes 6.2, 9.4, 10.8, 12.1, 14.1, 15.8, 16.3, 17.2, 19.0, 19.3, 20.1, 21.1, 21.7, 22.2, 22.7, 24.6, 25.4 and 29.2 degrees, and the measured 2θ value has an error of about ± 0.2 2θ.

[0086] In some embodiments, the powder X-ray diffraction characteristic diffraction angle (2θ) of Form AI of the Formula A salt includes 6.2, 9.4, 10.8, 12.1, 14.1, 15.8, 16.3, 17.2, 19.0, 19.3, 20.1, 21.1, 21.7, 22.2, 22.7, 24.6, 25.4, 26.8, 27.2, 27.5, 29.2, 30.6 and 31.8 degrees, and the measured 2θ value has an error of about ± 0.2 2θ.

[0087] In some embodiments, the crystalline form AI of the salt of formula A has Figure 1 The diffraction pattern shown. Despite experimental errors, instrument errors, and preferred orientation, a person skilled in the art can still obtain sufficient information to identify the crystal form AI of the salt of Formula A from the XRPD data provided herein.

[0088] In some embodiments, the crystalline form AI of the salt of formula A can be identified by differential scanning calorimetry. In some embodiments, the crystalline form AI of the salt of formula A has Figure 2 The differential scanning calorimetry analysis curve is shown in FIG. 1. In the DSC spectrum, the endothermic peaks of the crystal form AI of the salt of formula A are at about 162.8-179.6°C and 217.0-219.4°C.

[0089] In some embodiments, the crystalline form AI of the salt of formula A can be identified by thermogravimetric analysis. In some embodiments, the crystalline form AI of the salt of formula A has Figure 3The thermogravimetric analysis curve shown shows that there is a weight loss of about 10.96% between 100-170° C., which is the process of losing acetic acid due to heat. The crystal form AI is an anhydrate or pure crystal.

[0090] In some embodiments, the crystal form AI of the salt of formula A substantially does not contain other crystal forms described herein. For example, the weight content of the crystal form AI of the salt of formula A reaches at least 99%, at least 95%, at least 90%, or lower to 80%. Alternatively, the weight content of the crystal form AI of the salt of formula A reaches at least 70%, or at least 60%. Or further, the weight content of the crystal form AI of the salt of formula A reaches at least 50%.

[0091] Preparation method of crystal form AI

[0092] Method A

[0093] This article relates to a method for preparing a crystalline form AI of a salt of formula A, comprising:

[0094] (1) reacting a compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine with acetic acid to form a salt, and stirring the formed salt in at least one dissolving solvent or in a mixed solvent consisting of a water-miscible organic solvent and water;

[0095] (2) isolating and obtaining a crystalline solid of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine acetate;

[0096] (3) Optionally, drying the solid obtained in step (2).

[0097] In some embodiments, the molar ratio of acetic acid to compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine is no less than about 1:1. In some embodiments, the molar ratio is about 1.5:1. In some embodiments, the molar ratio is about 2:1. In some embodiments, the molar ratio is about 3:1. In some embodiments, the molar ratio is about 10:1. In some embodiments, the molar ratio is about 25:1.

[0098] In some embodiments, the ratio of the volume (ml) of the dissolving solvent or the mixed solvent in step (1) to the weight (g) of compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine is not less than about 10 ml / g (volume-to-mass ratio), for example, 13 ml / g, 18 ml / g, 23 ml / g, 24 ml / g, 25 ml / g, 31 ml / g, 35 ml / g, 90 ml / g, 100 ml / g.

[0099] In some embodiments, the dissolving solvent is selected from C 1-6 In some embodiments, the dissolving solvent is selected from the group consisting of methanol, ethanol, isopropanol, tert-butanol, dioxane, acetone and acetonitrile. In some embodiments, the dissolving solvent is selected from the group consisting of ethanol, isopropanol and dioxane.

[0100] In some embodiments, the water-miscible organic solvent accounts for less than about 95% by volume of the mixed solvent.

[0101] In some embodiments, the water-miscible organic solvent is selected from acetone, C 1-6 Alkyl alcohols (eg, methanol, ethanol, isopropanol), dioxane, and acetonitrile. In some embodiments, the water-miscible organic solvent is selected from acetone, ethanol, and dioxane.

[0102] In some embodiments, the water-miscible organic solvent and water are mixed in an appropriate ratio. In some embodiments, the volume ratio of the water-miscible organic solvent and water is from about 15:1 to 3:1, such as ethanol / water (volume ratio of about 11.6:1), acetone / water (volume ratio of about 11:1), dioxane / water (volume ratio of about 8:1).

[0103] In some embodiments, in step (1), the formation of the salt is carried out under heating and / or stirring, and the heating temperature should be no higher than the boiling point of the solvent system, such as about 40-50 degrees Celsius, about 60-70 degrees Celsius, and about 80-85 degrees Celsius.

[0104] In some embodiments, in step (1), the formed salt is stirred under cooling, preferably natural cooling, such as cooling to room temperature or lower temperature, such as about 20-25 degrees Celsius, about 5-10 degrees Celsius, and about 0-5 degrees Celsius.

[0105] In some embodiments, in step (1), the formed salt is stirred for 1-120 hours, such as at least 1 hour, at least 2 hours, at least 12 hours, at least 17 hours, at least 24 hours, at least 72 hours, at least 120 hours.

[0106] In some embodiments, in step (3), the drying temperature and drying time are appropriate so that the solid is fully dried and maintains the desired crystalline properties. In some embodiments, the drying temperature is 55 degrees Celsius. In some embodiments, the drying temperature is 60 degrees Celsius.

[0107] Method B

[0108] This article relates to another preparation method of the crystal form AI of the salt of formula A, comprising:

[0109] (1) adding compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine and acetic acid to an appropriate amount of at least one dissolving solvent or a mixed solvent consisting of a water-miscible organic solvent and water to react to form a salt, thereby obtaining a first solution;

[0110] (2) adding at least one anti-solvent to the first solution to obtain a second solution;

[0111] (3) isolating and obtaining a crystalline solid of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine acetate;

[0112] (4) Optionally, drying the solid obtained in step (3).

[0113] In some embodiments, the dissolving solvent is selected from C 1-6 Alkyl alcohols (eg ethanol, isopropanol) and dichloromethane.

[0114] In some embodiments, the water-miscible organic solvent is selected from C 1-6 Alkyl alcohol (eg, methanol, ethanol, isopropanol), dioxane, acetone and acetonitrile. In some embodiments, the water-miscible organic solvent is selected from ethanol, and the volume percentage of ethanol in the mixed solvent is not less than about 50%, such as 86.4%, 95%.

[0115] In some embodiments, the anti-dissolution solvent is selected from acetone, isopropyl ether, and methyl tert-butyl ether.

[0116] In some embodiments, the volume ratio of the dissolving solvent or mixed solvent to the anti-dissolving solvent is from about 1:2 to about 30:1, such as 1:2.3, 3.2:1, 4:1, 27.5:1.

[0117] In some embodiments, in step (1), stirring and heating may be performed simultaneously, and the heating temperature should be no higher than the boiling point of the solvent system, such as about 40-50 degrees Celsius, about 60-64 degrees Celsius, and about 80 degrees Celsius.

[0118] Crystal form BI

[0119] The present invention provides a crystalline form BI of a salt of formula A (wherein n is 1 and M is p-toluenesulfonic acid).

[0120] In some embodiments, the crystalline form BI of the salt of formula A can be identified by X-ray powder diffraction. In some embodiments, the powder X-ray diffraction characteristic diffraction angle (2θ) of the crystalline form BI of the salt of formula A includes 4.9, 5.5, 9.6, 14.4, 16.4 and 19.8 degrees, and the measured 2θ value has an error of about ± 0.2 2θ.

[0121] In some embodiments, the powder X-ray diffraction characteristic diffraction angles (2θ) of Form BI of the Formula A salt include 4.9, 5.5, 9.6, 14.4, 15.9, 16.4, 17.3, 18.4, 19.3 and 19.8 degrees, and the measured 2θ value has an error of about ±0.2 2θ.

[0122] In some embodiments, the powder X-ray diffraction characteristic diffraction angles (2θ) of Form BI include 4.9, 5.5, 6.8, 9.6, 10.1, 14.4, 14.8, 15.9, 16.4, 17.3, 18.4, 19.3, 19.8, 22.5 and 23.2 degrees, and the measured 2θ value has an error of about ± 0.2 2θ.

[0123] In some embodiments, the powder X-ray diffraction characteristic diffraction angle (2θ) of Form BI of the Formula A salt includes 4.9, 5.5, 6.8, 9.6, 10.1, 14.4, 14.8, 15.9, 16.4, 17.3, 18.4, 19.3, 19.8, 20.7, 21.4, 22.5, 23.2, 25.0, 26.1, 27.0 and 29.0 degrees, and the measured 2θ value has an error of about ± 0.2 2θ.

[0124] In some embodiments, the crystalline form BI of the salt of formula A has Figure 4The diffraction pattern shown. Despite experimental errors, instrument errors, and preferred orientation, a person skilled in the art can obtain sufficient information for identifying the crystalline form BI of the salt of Formula A from the XRPD data provided herein.

[0125] In some embodiments, the crystalline form BI of the salt of formula A can be identified by differential scanning calorimetry. In some embodiments, the crystalline form BI of the salt of formula A has Figure 5 The differential scanning calorimetry analysis curve shown is shown in FIG. 1. In the DSC spectrum, the endothermic peak of the crystal form BI of the salt of formula A is at about 205.7-209.6°C.

[0126] In some embodiments, the crystalline form BI of the salt of formula A can be identified by thermogravimetric analysis. In some embodiments, the crystalline form BI of the salt of formula A has Figure 6 Thermogravimetric analysis curve shown, which shows that the crystalline form BI is an anhydrate or pure crystal.

[0127] In some embodiments, the crystalline form BI of the salt of formula A substantially does not contain other crystalline forms described herein. For example, the weight content of the crystalline form BI of the salt of formula A reaches at least 99%, at least 95%, at least 90%, or lower to 80%. Alternatively, the weight content of the crystalline form BI of the salt of formula A reaches at least 70%, or at least 60%. Or further, the weight content of the crystalline form BI of the salt of formula A reaches at least 50%.

[0128] Preparation method of crystal form BI

[0129] Method A

[0130] This article relates to a method for preparing a crystalline form BI of a salt of formula A, comprising:

[0131] (1) reacting a compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine with p-toluenesulfonic acid to form a salt, and stirring the formed salt in a mixed solvent consisting of a water-miscible organic solvent and water;

[0132] (2) isolating and obtaining a crystalline solid of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine p-toluenesulfonate;

[0133] (3) Optionally, drying the solid obtained in step (2).

[0134] In some embodiments, the molar ratio of p-toluenesulfonic acid to compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine is no less than about 1: 1. In some embodiments, the molar ratio is about 1.1:1.

[0135] In some embodiments, the ratio of the volume (ml) of the mixed solvent in step (1) to the weight (g) of compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine is not less than about 10 ml / g (volume-to-mass ratio), for example, 25 ml / g, 32 ml / g, 33.5 ml / g, 37 ml / g, 42 ml / g, 52 ml / g.

[0136] In some embodiments, the volume percentage of the water-miscible organic solvent in the mixed solvent is less than about 96%.

[0137] In some embodiments, the water-miscible organic solvent is selected from C 1-6 Alkyl alcohols (eg, ethanol, isopropanol) and acetone. In some embodiments, the water-miscible organic solvent is selected from ethanol and isopropanol.

[0138] In some embodiments, the water-miscible organic solvent and water are mixed in an appropriate ratio. In some embodiments, the volume ratio of the water-miscible organic solvent and water is from about 22:1 to 4:1, such as ethanol / water (volume ratio of about 4.3:1, about 7.7:1 or about 13:1), isopropanol / water (volume ratio of about 4.3:1 or about 17.4:1), acetone / water (volume ratio of about 22:1).

[0139] In some embodiments, in step (1), the formation of the salt is carried out under heating and / or stirring, and the heating temperature should not be higher than the boiling point of the solvent system, such as about 50-60 degrees Celsius and about 80-90 degrees Celsius.

[0140] In some embodiments, in step (1), the formed salt is stirred under cooling, preferably natural cooling, such as cooling to room temperature or lower temperature, such as about 15-20 degrees Celsius.

[0141] In some embodiments, in step (1), the formed salt is stirred for 1-120 hours, such as at least 3 hours, at least 16 hours, or at least 72 hours.

[0142] In some embodiments, in step (3), the drying temperature and drying time are appropriate so that the solid is fully dried and maintains the desired crystalline properties. In some embodiments, the drying temperature is room temperature. In some embodiments, the drying temperature is 60 degrees Celsius.

[0143] Method B

[0144] This invention relates to another method for preparing the crystalline form BI of the salt of formula A, comprising:

[0145] The crystalline form B-II of the salt of formula A is stirred in about 95% ethanol at a temperature of about 10-30° C., such as at room temperature, to obtain the crystalline form BI.

[0146] In some embodiments, the stirring period is 4 days.

[0147] Form B-II

[0148] The present invention provides a crystalline form B-II of a salt of formula A (wherein n is 1 and M is p-toluenesulfonic acid).

[0149] In some embodiments, the crystalline form B-II of the salt of formula A can be identified by X-ray powder diffraction. In some embodiments, the powder X-ray diffraction characteristic diffraction angle (2θ) of the crystalline form B-II of the salt of formula A includes 5.1, 6.0, 10.2, 17.1 and 19.1 degrees, and the measured 2θ value has an error of about ± 0.2 2θ.

[0150] In some embodiments, the powder X-ray diffraction characteristic diffraction angles (2θ) of Form B-II of the salt of Formula A include 5.1, 6.0, 9.5, 10.2, 14.8, 15.8, 17.1, 19.1 and 22.4 degrees, and the measured 2θ value has an error of about ±0.2 2θ.

[0151] In some embodiments, the powder X-ray diffraction characteristic diffraction angles (2θ) of Form B-II include 5.1, 6.0, 9.5, 10.2, 14.3, 14.8, 15.8, 17.1, 19.1, 20.2, 20.8, 22.4 and 26.0 degrees, and the measured 2θ value has an error of about ±0.22θ.

[0152] In some embodiments, the powder X-ray diffraction characteristic diffraction angle (2θ) of Form B-II of the salt of Formula A includes 5.1, 6.0, 9.5, 10.2, 14.3, 14.8, 15.3, 15.8, 17.1, 17.9, 19.1, 19.7, 20.2, 20.8, 22.4, 23.4, 26.0 and 27.2 degrees, and the measured 2θ value has an error of about ±0.2 2θ.

[0153] In some embodiments, the crystalline form B-II of the salt of formula A has Figure 7 The diffraction pattern shown. Despite experimental errors, instrument errors, and preferred orientation, a person skilled in the art can still obtain sufficient information to identify the crystalline form B-II of the salt of Formula A from the XRPD data provided herein.

[0154] In some embodiments, the crystalline form B-II of the salt of formula A can be identified by differential scanning calorimetry. In some embodiments, the crystalline form B-II of the salt of formula A has Figure 8 The differential scanning calorimetry analysis curve shown in FIG. 1 shows that in the DSC spectrum, the endothermic peak of the crystalline form B-II of the salt of formula A is at about 203.0-211.4°C.

[0155] In some embodiments, the crystalline form B-II of the salt of formula A can be identified by thermogravimetric analysis. In some embodiments, the crystalline form B-II of the salt of formula A has Figure 8 Thermogravimetric analysis curves shown show that the crystalline form B-II is an anhydrate or pure crystal.

[0156] In some embodiments, the crystalline form B-II of the salt of formula A is substantially free of other crystalline forms described herein. For example, the weight content of the crystalline form B-II of the salt of formula A reaches at least 99%, at least 95%, at least 90%, or lower to 80%. Alternatively, the weight content of the crystalline form B-II of the salt of formula A reaches at least 70%, or at least 60%. Or further, the weight content of the crystalline form B-II of the salt of formula A reaches at least 50%.

[0157] Preparation method of crystal form B-II

[0158] This article relates to a method for preparing the crystalline form B-II of the salt of formula A, comprising:

[0159] (1) reacting a compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine with p-toluenesulfonic acid to form a salt, and stirring the formed salt in a mixed solvent consisting of methanol and water;

[0160] (2) isolating and obtaining a crystalline solid of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine p-toluenesulfonate;

[0161] (3) Optionally, drying the solid obtained in step (2).

[0162] In some embodiments, the molar ratio of p-toluenesulfonic acid to compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine is no less than about 1: 1. In some embodiments, the molar ratio is about 1.1:1.

[0163] In some embodiments, the ratio of the volume (ml) of the mixed solvent in step (1) to the weight (g) of compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine is not less than about 10 ml / g (volume-to-mass ratio), for example, about 32 ml / g.

[0164] In some embodiments, the volume ratio of methanol to water in step (1) is about 13:1.

[0165] In some embodiments, in step (1), the formation of the salt is carried out under heating and / or stirring, and the heating temperature should not be higher than the boiling point of the solvent system.

[0166] In some embodiments, in step (1), the formed salt is stirred under cooling, preferably natural cooling, for example, cooling to room temperature.

[0167] In some embodiments, in step (1), the formed salt is stirred for 1-120 hours, for example, at least 72 hours.

[0168] Hydrate (Form B-III)

[0169] The present invention also provides a hydrate of the salt of formula A (wherein n is 1 and M is p-toluenesulfonic acid).

[0170] In some embodiments, the hydrate of the salt of Formula A (wherein n is 1 and M is p-toluenesulfonic acid) is a hemihydrate.

[0171] In some embodiments, the hemihydrate of the salt of Formula A (wherein n is 1 and M is p-toluenesulfonic acid) is Form B-III.

[0172] In some embodiments, the crystalline form B-III of the hemihydrate of the salt of formula A can be identified by X-ray powder diffraction. The powder X-ray diffraction characteristic diffraction angles (2θ) of the crystalline form B-III include 5.3, 5.9, 10.7, 13.6 and 15.6 degrees, and the measured 2θ value has an error of about ±0.2 2θ.

[0173] In some embodiments, the powder X-ray diffraction characteristic diffraction angles (2θ) of Form B-III of the hemihydrate of the salt of Formula A include 5.3, 5.9, 9.9, 10.7, 11.8, 13.6, 14.9, 15.6 and 17.6 degrees, and the measured 2θ value has an error of about ±0.2 2θ.

[0174] In some embodiments, the powder X-ray diffraction characteristic diffraction angles (2θ) of Form B-III of the hemihydrate of the salt of Formula A include 5.3, 5.9, 9.9, 10.7, 11.8, 13.6, 14.9, 15.6, 16.0, 17.6, 20.0 and 22.8 degrees, and the measured 2θ value has an error of about ±0.2 2θ.

[0175] In some embodiments, the powder X-ray diffraction characteristic diffraction angle (2θ) of Form B-III of the hemihydrate of the salt of Formula A includes 5.3, 5.9, 9.9, 10.7, 11.8, 13.6, 14.9, 15.6, 16.0, 17.6, 18.9, 20.0, 21.6, 22.8, 25.0 and 27.0 degrees, and the measured 2θ value has an error of about ± 0.2 2θ.

[0176] In some embodiments, the crystalline form B-III of the hemihydrate of the salt of formula A has Fig. 9 The diffraction pattern shown. Despite experimental errors, instrument errors, and preferred orientation, a person skilled in the art can obtain sufficient information to identify the hemihydrate of the salt of Formula A as Form B-III from the XRPD data provided herein.

[0177] In some embodiments, the crystalline form B-III of the hemihydrate of the salt of formula A can be identified by differential scanning calorimetry. In some embodiments, the crystalline form B-III of the hemihydrate of the salt of formula A has Fig.10 The differential scanning calorimetry analysis curve is shown in FIG. 1. In the DSC spectrum, the hemihydrate of the salt of Formula A, Form B-III, has an exothermic peak at about 168.1-172.5°C and an obvious endothermic peak at about 199.6-208.3°C.

[0178] In some embodiments, the crystalline form B-III of the hemihydrate of the salt of formula A can be identified by thermogravimetric analysis. In some embodiments, the crystalline form B-III of the hemihydrate of the salt of formula A has Fig.10 The thermogravimetric analysis curve shown in the figure shows that the crystal form B-III is a hemihydrate.

[0179] In some embodiments, the crystalline form B-III of the hemihydrate of the salt of formula A is substantially free of other crystalline forms described herein. For example, the weight content of the crystalline form B-III of the hemihydrate of the salt of formula A reaches at least 99%, at least 95%, at least 90%, or lower to 80%. Alternatively, the weight content of the crystalline form B-III of the hemihydrate of the salt of formula A reaches at least 70%, or at least 60%. Or further, the weight content of the crystalline form B-III of the hemihydrate of the salt of formula A reaches at least 50%.

[0180] Preparation method of crystal form B-III

[0181] Method A

[0182] The present invention relates to a method for preparing a crystalline form B-III of a hydrate of a salt of formula A, comprising:

[0183] (1) reacting a compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine with p-toluenesulfonic acid to form a salt, and stirring the formed salt in water;

[0184] (2) isolating and obtaining a crystalline solid of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine p-toluenesulfonate;

[0185] (3) Optionally, drying the solid obtained in step (2).

[0186] In some embodiments, the molar ratio of p-toluenesulfonic acid to compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine is no less than about 1: 1. In some embodiments, the molar ratio is about 1.1:1.

[0187] In some embodiments, the ratio of the volume (ml) of water in step (1) to the weight (g) of compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine is about 54 ml / g (volume-to-mass ratio).

[0188] In some embodiments, in step (1), the formation of the salt is carried out under heating and / or stirring, and the heating temperature should be appropriate, for example, about 80-85 degrees Celsius.

[0189] In some embodiments, in step (1), the formed salt is stirred under cooling, preferably natural cooling, for example, cooling to room temperature.

[0190] Method B

[0191] The present invention relates to another method for preparing the crystalline form B-III of the hydrate of the salt of formula A, comprising:

[0192] The crystalline form BI of the salt of formula A is stirred in water at a temperature of about 10-30° C., for example at room temperature, and the solid is collected by filtration and optionally dried to obtain the crystalline form B-III.

[0193] In some embodiments, the stirring period is 4 days.

[0194] In some embodiments, the drying is vacuum drying. In some embodiments, the drying temperature and drying time are appropriate so that the solid is fully dried and maintains the desired crystalline properties. In some embodiments, the drying temperature is 60 degrees Celsius. In some embodiments, the drying time is 2 hours.

[0195] Crystal form CI

[0196] The present invention provides a crystal form CI of a salt of formula A (wherein n is 1 and M is malic acid).

[0197] In some embodiments, the crystal form CI of the salt of formula A can be identified by X-ray powder diffraction. In some embodiments, the powder X-ray diffraction characteristic diffraction angle (2θ) of the crystal form CI of the salt of formula A includes 8.6, 14.3, 15.5, 19.5 and 22.2 degrees, and the measured 2θ value has an error of about ± 0.2 2θ.

[0198] In some embodiments, the powder X-ray diffraction characteristic diffraction angles (2θ) of Form CI of Formula A salt include 8.6, 10.8, 14.3, 15.5, 16.4, 17.7, 18.4, 19.5, 22.2 and 23.8 degrees, and the measured 2θ value has an error of about ±0.2 2θ.

[0199] In some embodiments, the powder X-ray diffraction characteristic diffraction angles (2θ) of Form CI include 8.6, 10.8, 14.3, 15.5, 16.4, 17.2, 17.7, 18.4, 19.5, 20.9, 22.2, 22.6, 23.8, 29.2, 29.8 and 30.7 degrees, and the measured 2θ value has an error of about ±0.2 2θ.

[0200] In some embodiments, the powder X-ray diffraction characteristic diffraction angle (2θ) of Form CI of the Formula A salt includes 8.6, 10.8, 11.8, 13.5, 14.3, 15.5, 16.4, 17.2, 17.7, 18.1, 18.4, 19.5, 20.9, 22.2, 22.6, 23.8, 25.8, 26.7, 27.8, 29.2, 29.8 and 30.7 degrees, and the measured 2θ value has an error of about ± 0.2 2θ.

[0201] In some embodiments, the crystalline form CI of the salt of Formula A has Fig.11 The diffraction pattern shown. Despite experimental errors, instrument errors, and preferred orientation, a person skilled in the art can still obtain sufficient information to identify the crystal form CI of the salt of Formula A from the XRPD data provided herein.

[0202] In some embodiments, the crystalline form CI of the salt of formula A can be identified by differential scanning calorimetry. In some embodiments, the crystalline form CI of the salt of formula A has Fig.12 The differential scanning calorimetry analysis curve is shown in FIG. 1. In the DSC spectrum, the endothermic peak of the crystal form CI of the salt of formula A is at about 211.1-214.9°C.

[0203] In some embodiments, the crystalline form CI of the salt of formula A can be identified by thermogravimetric analysis. In some embodiments, the crystalline form CI of the salt of formula A has Fig.13 Thermogravimetric analysis curves shown show that the crystal form CI is an anhydrate or pure crystal.

[0204] In some embodiments, the crystal form CI of the salt of formula A is substantially free of other crystal forms described herein. For example, the weight content of the crystal form CI of the salt of formula A reaches at least 99%, at least 95%, at least 90%, or lower to 80%. Alternatively, the weight content of the crystal form CI of the salt of formula A reaches at least 70%, or at least 60%. Or further, the weight content of the crystal form CI of the salt of formula A reaches at least 50%.

[0205] Preparation method of crystal form CI

[0206] This article relates to a method for preparing a crystalline form CI of a salt of formula A, comprising:

[0207] (1) reacting a compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine with malic acid to form a salt, and stirring the formed salt in at least one dissolving solvent or in a mixed solvent consisting of a water-miscible organic solvent and water;

[0208] (2) isolating and obtaining a crystalline CI solid of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine malate;

[0209] (3) Optionally, drying the solid obtained in step (2).

[0210] In some embodiments, the molar ratio of malic acid to compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine is no less than about 1:1. In some embodiments, the molar ratio is about 1.1:1. In some embodiments, the molar ratio is about 1.5:1.

[0211] In some embodiments, the ratio of the volume (ml) of the dissolving solvent or the mixed solvent in step (1) to the weight (g) of compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine is not less than about 30 ml / g (volume-to-mass ratio), for example, 38 ml / g, 53 ml / g, 67 ml / g, 86 ml / g, or 135 ml / g.

[0212] In some embodiments, the dissolving solvent is selected from C 1-6 Alkyl alcohols (eg, methanol, ethanol, isopropanol) and tetrahydrofuran. In some embodiments, the dissolving solvent is selected from ethanol.

[0213] In some embodiments, the water-miscible organic solvent is selected from C 1-6 Alkyl alcohols (eg methanol, ethanol, isopropanol) and tetrahydrofuran.

[0214] In some embodiments, the water-miscible organic solvent and water are mixed in an appropriate ratio. In some embodiments, the volume ratio of the water-miscible organic solvent and water is from about 12:1 to 1:1, such as ethanol / water (volume ratio of about 1.5:1), methanol / water (volume ratio of about 1.5:1), isopropanol / water (volume ratio of about 1.4:1), tetrahydrofuran / water (volume ratio of about 12:1).

[0215] In some embodiments, in step (1), the formation of the salt is carried out under heating and / or stirring, and the heating temperature should be no higher than the boiling point of the solvent system, such as about 40-50 degrees Celsius, about 60-70 degrees Celsius, and about 75-85 degrees Celsius.

[0216] In some embodiments, in step (1), the formed salt is stirred under cooling, preferably natural cooling, such as cooling to room temperature or lower temperature, such as about 15-20 degrees Celsius.

[0217] In some embodiments, in step (1), the formed salt is stirred for 1-120 hours, such as at least 12 hours, at least 14 hours, at least 20 hours, at least 72 hours.

[0218] In some embodiments, in step (3), the drying temperature and drying time should be appropriate so that the solid is fully dried and maintains the desired crystal properties. In some embodiments, the drying temperature is 60 degrees Celsius.

[0219] Hydrate (Crystal Form C-II)

[0220] The present invention also provides a hydrate of the salt of formula A (wherein n is 0.5 and M is malic acid).

[0221] In some embodiments, the hydrate of the salt of formula A (wherein n is 0.5 and M is malic acid) contains 0.75 molecules of water.

[0222] In some embodiments, the hydrate of the salt of formula A (wherein n is 0.5 and M is malic acid) containing 0.75 molecules of water is Form C-II.

[0223] In some embodiments, the hydrated form C-II of the salt of Formula A can be identified by X-ray powder diffraction. The powder X-ray diffraction characteristic diffraction angles (2θ) of the crystalline form C-II include 11.3, 11.6, 17.0, 20.0, 21.6 and 23.1 degrees, and the measured 2θ value has an error of about ±0.2 2θ.

[0224] In some embodiments, the powder X-ray diffraction characteristic diffraction angles (2θ) of Form C-II of the hydrate of the salt of Formula A include 5.3, 9.5, 10.0, 11.3, 11.6, 17.0, 20.0, 21.6, 23.1, 26.9 and 28.3 degrees, and the measured 2θ value has an error of about ±0.2 2θ.

[0225] In some embodiments, the powder X-ray diffraction characteristic diffraction angle (2θ) of Form C-II of the hydrate of the salt of Formula A includes 5.3, 9.5, 10.0, 10.6, 11.3, 11.6, 12.4, 12.8, 17.0, 20.0, 21.6, 23.1, 24.9, 26.4, 26.9 and 28.3 degrees, and the measured 2θ value has an error of about ±0.2 2θ.

[0226] In some embodiments, the powder X-ray diffraction characteristic diffraction angle (2θ) of Form C-II of the hydrate of the salt of Formula A includes 5.3, 9.5, 10.0, 10.6, 11.3, 11.6, 12.4, 12.8, 13.6, 15.9, 17.0, 18.4, 20.0, 20.9, 21.6, 23.1, 24.9, 26.4, 26.9 and 28.3 degrees, and the measured 2θ value has an error of about ±0.22θ.

[0227] In some embodiments, the crystalline form C-II of the hydrate of the salt of formula A has Fig.14 The diffraction pattern shown. Despite experimental errors, instrument errors, and preferred orientation, a person skilled in the art can obtain sufficient information to identify the hydrated form C-II of the salt of Formula A from the XRPD data provided herein.

[0228] In some embodiments, the crystalline form C-II of the hydrate of the salt of formula A can be identified by differential scanning calorimetry. In some embodiments, the crystalline form C-II of the hydrate of the salt of formula A has Fig.15 The differential scanning calorimetry analysis curve is shown in FIG. 1. In the DSC spectrum, the endothermic peaks of the hydrated form C-II of the salt of Formula A are at about 70.2-90.4°C and about 202.6-215.0°C.

[0229] In some embodiments, the crystalline form C-II of the hydrate of the salt of formula A can be identified by thermogravimetric analysis. In some embodiments, the crystalline form C-II of the hydrate of the salt of formula A has Fig.16 The thermogravimetric analysis curve shown in the figure shows that the weight loss is 2.61% near 30-90°C, and the crystal form C-II is a hydrate containing 0.75 molecules of water.

[0230] In some embodiments, the crystalline form C-II of the hydrate of the salt of formula A is substantially free of other crystalline forms described herein. For example, the weight content of the crystalline form C-II of the hydrate of the salt of formula A reaches at least 99%, at least 95%, at least 90%, or lower to 80%. Alternatively, the weight content of the crystalline form C-II of the hydrate of the salt of formula A reaches at least 70%, or at least 60%. Or further, the weight content of the crystalline form C-II of the hydrate of the salt of formula A reaches at least 50%.

[0231] Preparation method of crystal form C-II

[0232] The present invention relates to a method for preparing a crystalline form C-II of a hydrate of a salt of formula A, comprising:

[0233] (1) reacting a compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine with malic acid to form a salt, and stirring the formed salt in a mixed solvent consisting of acetonitrile and water;

[0234] (2) isolating and obtaining a crystalline solid of Form C-II of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine malate;

[0235] (3) Optionally, drying the solid obtained in step (2).

[0236] In some embodiments, the molar ratio of malic acid to compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine is no less than about 1: 1. In some embodiments, the molar ratio is about 1.1:1.

[0237] In some embodiments, the ratio of the volume (ml) of the mixed solvent in step (1) to the weight (g) of compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine is about 88 ml / g (volume-to-mass ratio).

[0238] In some embodiments, the volume ratio of water to acetonitrile in step (1) is not less than about 6: 1. In some embodiments, the volume ratio is about 6:1.

[0239] In some embodiments, in step (1), the formation of the salt is carried out under heating and / or stirring, and the heating temperature should not be higher than the boiling point of the solvent system, for example, about 80-85 degrees Celsius.

[0240] In some embodiments, in step (1), the formed salt is stirred under cooling, preferably natural cooling, for example, cooling to room temperature.

[0241] In some embodiments, in step (1), the formed salt is stirred for 1-120 hours, for example, at least 18 hours.

[0242] Hydrate (Form C-III)

[0243] The present invention also provides a hydrate of the salt of formula A (wherein n is 0.5 and M is malic acid).

[0244] In some embodiments, the hydrate of the salt of Formula A (wherein n is 0.5 and M is malic acid) is a monohydrate.

[0245] In some embodiments, the monohydrate of the salt of Formula A (wherein n is 0.5 and M is malic acid) is Form C-III.

[0246] In some embodiments, the monohydrate of the salt of Formula A in Form C-III can be identified by X-ray powder diffraction. The powder X-ray diffraction characteristic diffraction angles (2θ) of Form C-III include 5.3, 10.6, 17.0, 17.9 and 25.2 degrees, and the measured 2θ value has an error of about ±0.2 2θ.

[0247] In some embodiments, the powder X-ray diffraction characteristic diffraction angles (2θ) of Form C-III of the monohydrate of the salt of Formula A include 5.3, 10.6, 12.8, 17.0, 17.9, 20.3, 21.7, 22.5 and 25.2 degrees, and the measured 2θ value has an error of about ±0.2 2θ.

[0248] In some embodiments, the powder X-ray diffraction characteristic diffraction angles (2θ) of Form C-III of the monohydrate of the salt of Formula A include 5.3, 10.6, 11.8, 12.8, 16.0, 17.0, 17.9, 18.9, 20.3, 21.7, 22.5 and 25.2 degrees, and the measured 2θ value has an error of about ±0.2 2θ.

[0249] In some embodiments, the powder X-ray diffraction characteristic diffraction angle (2θ) of Form C-III of the monohydrate of the salt of Formula A includes 5.3, 10.6, 11.8, 12.8, 16.0, 17.0, 17.9, 18.6, 18.9, 20.3, 21.7, 22.5, 23.1, 25.2, 25.6 and 27.4 degrees, and the measured 2θ value has an error of about ± 0.2 2θ.

[0250] In some embodiments, the crystalline form C-III of the monohydrate of the salt of Formula A has Fig.17 The diffraction pattern shown. Despite experimental errors, instrument errors, and preferred orientation, a person skilled in the art can obtain sufficient information to identify the monohydrate form C-III of the salt of Formula A from the XRPD data provided herein.

[0251] In some embodiments, the crystalline form C-III of the monohydrate of the salt of formula A can be identified by differential scanning calorimetry. In some embodiments, the crystalline form C-III of the monohydrate of the salt of formula A has Fig.18 The differential scanning calorimetry analysis curve is shown in FIG. 1. In the DSC spectrum, the endothermic peaks of the monohydrate form C-III of the salt of Formula A are at about 69.9-88.5°C and about 201.6-206.7°C.

[0252] In some embodiments, the crystalline form C-III of the monohydrate of the salt of formula A can be identified by thermogravimetric analysis. In some embodiments, the crystalline form C-III of the monohydrate of the salt of formula A has Fig.19 The thermogravimetric analysis curve shown in the figure shows that the weight loss is 3.2% near 30-90°C, and the crystal form C-III is a monohydrate.

[0253] In some embodiments, the crystalline form C-III of the monohydrate of the salt of formula A is substantially free of other crystalline forms described herein. For example, the weight content of the crystalline form C-III of the monohydrate of the salt of formula A reaches at least 99%, at least 95%, at least 90%, or lower to 80%. Alternatively, the weight content of the crystalline form C-III of the monohydrate of the salt of formula A reaches at least 70%, or at least 60%. Or further, the weight content of the crystalline form C-III of the monohydrate of the salt of formula A reaches at least 50%.

[0254] Preparation method of crystal form C-III

[0255] The present invention relates to a method for preparing a crystalline form C-III of a hydrate of a salt of formula A, comprising:

[0256] (1) reacting a compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine with malic acid to form a salt, and stirring the formed salt in a mixed solvent consisting of ethanol and water;

[0257] (2) isolating and obtaining a crystalline solid of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine malate;

[0258] (3) Optionally, drying the solid obtained in step (2).

[0259] In some embodiments, the molar ratio of malic acid to compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine is no less than about 1: 1. In some embodiments, the molar ratio is about 1.5:1.

[0260] In some embodiments, the ratio of the volume (ml) of the mixed solvent in step (1) to the weight (g) of compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine is about 55 ml / g (volume-to-mass ratio).

[0261] In some embodiments, the volume ratio of water to ethanol in step (1) is not less than about 4: 1. In some embodiments, the volume ratio is about 4:1.

[0262] In some embodiments, in step (1), the formation of the salt is carried out under heating and / or stirring, and the heating temperature should not be higher than the boiling point of the solvent system, for example, about 80 degrees Celsius.

[0263] In some embodiments, in step (1), the formed salt is stirred under cooling, preferably natural cooling, such as cooling to room temperature or lower temperature, such as about 15-20 degrees Celsius.

[0264] In some embodiments, in step (1), the formed salt is stirred for 1-120 hours, for example, at least 12 hours.

[0265] The features in the various embodiments of the above-mentioned preparation methods of each crystalline form of the salt of Formula A or its solvate can be arbitrarily combined with each other, and the various schemes obtained by these combinations are included in the scope of the present invention, just as the schemes obtained by these combinations are specifically and one by one listed in this document.

[0266] Pharmaceutical compositions and methods of treatment

[0267] The salt of formula A or its solvate or the crystalline form of the salt of formula A or its solvate (e.g., crystalline form AI, crystalline form BI, crystalline form B-II, crystalline form B-III, crystalline form CI, crystalline form C-II and crystalline form C-III) can be used to treat diseases, such as autoimmune diseases, inflammatory diseases and cancer. The cancer is preferably a hematological malignancy. The autoimmune disease, inflammatory disease and cancer include but are not limited to systemic lupus erythematosus, rheumatoid arthritis, Sjögren's syndrome, IgA nephropathy, autoimmune hemolytic anemia, idiopathic thrombocytopenic purpura, multiple sclerosis, myasthenia gravis, allergic rhinitis, chronic obstructive pulmonary disease, adult respiratory distress syndrome, asthma, lymphoma (e.g., B cell lymphoma, T cell lymphoma), leukemia (e.g., chronic lymphocytic leukemia, acute lymphocytic leukemia and acute myeloid leukemia) and multiple myeloma.

[0268] Provided herein is a method for treating a disease responsive to inhibition of Syk kinase activity, comprising administering an active pharmaceutical ingredient formed from a salt of formula A, or a salt of formula A or a solvate thereof of the present invention, or a crystalline form of a salt of formula A or a solvate thereof, such as one or more of crystalline form AI, crystalline form BI, crystalline form B-II, crystalline form B-III, crystalline form CI, crystalline form C-II, or crystalline form C-III.

[0269] In some embodiments, this treatment method is directed to at least one disease that responds to inhibition of Syk kinase activity, such as autoimmune diseases, inflammatory diseases, and cancer (preferably hematological malignancies). Wherein, an effective amount of the pharmaceutical composition of the present invention is administered to an individual in need for treatment, wherein the pharmaceutical composition comprises at least one pharmaceutically acceptable carrier, and one or more of a salt of formula A or a solvate thereof or a crystalline form of a salt of formula A or a solvate thereof (e.g., crystalline form AI, crystalline form BI, crystalline form B-II, crystalline form B-III, crystalline form CI, crystalline form C-II, or crystalline form C-III).

[0270] The dosage of at least one active pharmaceutical ingredient selected from a salt of formula A, or a crystalline form of a salt of formula A, or a solvate of a salt of formula A or a crystalline form thereof (e.g., Form AI, Form BI, Form B-II, Form B-III, Form CI, Form C-II, or Form C-III) to achieve the desired physiological effect depends on a variety of factors, such as the purpose of use, the mode of administration, and the clinical condition of the patient. The daily dose may be, for example, ranging from 0.01 mg to 3 g per day (e.g., from 0.05 mg to 2 g per day, or even from 100 mg to 1 g per day). Orally administrable unit dosage formulations include, for example, tablets or capsules.

[0271] To achieve the above-mentioned therapeutic purposes, at least one active pharmaceutical ingredient selected from the salt of formula A or its solvate or the crystalline form of the salt of formula A or its solvate can be administered in the form of the compound itself, but usually they are used in the form of a pharmaceutical composition composed of one or more pharmaceutically acceptable carriers or excipients.

[0272] Representative carriers or excipients should be compatible with other ingredients in the composition and will not endanger the health of the patient. The carrier or excipient can be solid or liquid, or both, and they form a pharmaceutical composition or unit dosage form (e.g., tablets, capsules) with the salt of formula A or its solvate or the crystalline form of the salt of formula A or its solvate (e.g., crystalline form AI, crystalline form BI, crystalline form B-II, crystalline form B-III, crystalline form CI, crystalline form C-II and / or crystalline form C-III), which can contain 0.05% to 95% of the salt of formula A by weight. The pharmaceutical composition described in the present invention can be prepared by known drug preparation methods, such as the method including mixing with pharmaceutically acceptable carriers and / or excipients and diluents.

[0273] In some examples, at least one active pharmaceutical ingredient selected from a salt of formula A or a solvate thereof or a crystalline form of a salt of formula A or a solvate thereof (e.g., Form AI, Form BI, Form B-II, Form B-III, Form CI, Form C-II, and Form C-III) may be combined with at least one component, such as a carrier and / or an excipient and / or a diluent, which may be selected from sweeteners, flavoring agents, colorants, dyes, and emulsifiers.

[0274] In some examples, the crystal form of the salt of formula A or its solvate or the crystal form of the salt of formula A or its solvate (e.g., crystal form AI, crystal form BI, crystal form B-II, crystal form B-III, crystal form CI, crystal form C-II, and crystal form C-III) will not be transformed when it is made into a preparation with one or more pharmaceutically acceptable carriers and / or excipients and / or diluents. In other examples, the crystal form of the salt of formula A or its solvate or the crystal form of the salt of formula A or its solvate (e.g., crystal form AI, crystal form BI, crystal form B-II, crystal form B-III, crystal form CI, crystal form C-II, or crystal form C-III) may be transformed when it is made into a preparation with one or more pharmaceutically acceptable carriers and / or excipients and / or diluents, and it may be completely or partially transformed into one or more crystal forms, including transformation into a non-solid form. In some examples, the crystal form AI or other crystal forms of the present invention can be dissolved when the pharmaceutical composition is made. Therefore, in these "dissolved" examples, the crystal form AI or other crystal forms no longer exist in their respective crystal forms in the pharmaceutical composition.

[0275] In some examples, at least one active pharmaceutical ingredient selected from a salt of formula A or a solvate thereof or a crystalline form of a salt of formula A or a solvate thereof (e.g., Form AI, Form BI, Form B-II, Form B-III, Form CI, Form C-II, and Form C-III) is formulated into a suitable preparation for administration.

[0276] The pharmaceutical compositions described in the present invention may be those in dosage forms suitable for oral and oral (such as sublingual) administration, and the appropriate administration method may depend on the condition of each case and the severity of the treatment, and also on the properties of the specific form of the active pharmaceutical ingredient selected from the salt of formula A or its solvate or the crystalline form of the salt of formula A or its solvate (e.g., crystalline form AI, crystalline form BI, crystalline form B-II, crystalline form B-III, crystalline form CI, crystalline form C-II and crystalline form C-III) used in the preparation of the pharmaceutical composition.

[0277] A suitable pharmaceutical composition for oral administration prepared from at least one active pharmaceutical ingredient selected from a salt of formula A or a solvate thereof or a crystalline form of a salt of formula A or a solvate thereof (e.g., crystalline form AI, crystalline form BI, crystalline form B-II, crystalline form B-III, crystalline form CI, crystalline form C-II and crystalline form C-III) may also be in the form of a unit dosage form, for example, capsules, cachets and tablets, including suckable tablets, each of which is quantitatively prepared from at least one active pharmaceutical ingredient described in the present invention; its preparation form may also be selected from powders, granules, solutions, suspensions in water or non-aqueous liquids, oil-in-water emulsions and water-in-oil emulsions. These compositions can also be prepared by any suitable method for preparing pharmaceutical preparations as described above, for example, these methods include the following steps: mixing at least one active pharmaceutical ingredient selected from the salt of formula A or its solvate or the crystalline form of the salt of formula A or its solvate (e.g., crystalline form AI, crystalline form BI, crystalline form B-II, crystalline form B-III, crystalline form CI, crystalline form C-II and crystalline form C-III) and a carrier and / or excipient and / or diluent (which may be composed of one or more additional ingredients). These compositions can usually be prepared by uniformly and homogeneously mixing at least one active pharmaceutical ingredient selected from the salt of formula A or its solvate or the crystalline form of the salt of formula A or its solvate (e.g., crystalline form AI, crystalline form BI, crystalline form B-II, crystalline form B-III, crystalline form CI, crystalline form C-II and crystalline form C-III) and a liquid or finely divided solid carrier, and the product can be made into a certain shape.

[0278] At least one active pharmaceutical ingredient selected from a salt of formula A or a solvate thereof or a crystalline form of a salt of formula A or a solvate thereof (e.g., Form AI, Form BI, Form B-II, Form B-III, Form CI, Form C-II, and Form C-III) can also be used in combination with one or more other active ingredients (e.g., in collaborative therapy). When used in combination, the active ingredients can be separate compositions for simultaneous administration or respectively (e.g., sequential administration in any order) at different times in treatment by the same or different routes of administration, or they can also be administered together in the same pharmaceutical composition.

[0279] In some examples, at least one active pharmaceutical ingredient selected from a salt of formula A or a solvate thereof or a crystalline form of a salt of formula A or a solvate thereof (e.g., Form AI, Form BI, Form B-II, Form B-III, Form CI, Form C-II, and Form C-III) can be administered simultaneously with one or more other active ingredients known to have therapeutic effects, such as for the treatment of diseases responsive to inhibition of Syk kinase activity, such as autoimmune diseases, inflammatory diseases, and cancers (preferably hematological malignancies).

[0280] The term "combination use" as used herein is used to define the combination of at least one active pharmaceutical ingredient selected from the salt of Formula A or its solvate or the crystalline form of the salt of Formula A or its solvate (e.g., crystalline form AI, crystalline form BI, crystalline form B-II, crystalline form B-III, crystalline form CI, crystalline form C-II and crystalline form C-III) with one or more other active ingredients, such as in the treatment of autoimmune diseases or inflammatory diseases (e.g., in combination with anti-immune agents, steroids), in anti-tumor methods (e.g., in combination with BTK inhibitors, PI3Kδ inhibitors, Bcl-2 inhibitors, lenalidomide). Examples of BTK inhibitors include, but are not limited to, ibrutinib, ACP-196 (Acalabrutinib), CC-292 (Spebrutinib), ONO-4059 (Tirabrutinib), BGB-3111 and GDC-0853. Examples of PI3Kδ inhibitors include, but are not limited to, Idelalisib, IPI-145 (Duvelisib), TGR-1202 (Umbralisib), GS-9820 (Acalisib), and INCB-050465. Examples of Bcl-2 inhibitors include, but are not limited to, Venetoclax (ABT-199) and ABT-263 (Navitoclax). Here, "anti-tumor method" may refer to any method for the purpose of treating cancer (including hematological malignancies). Examples of anti-tumor methods include, but are not limited to, chemotherapy, radiotherapy, targeted therapy, and immunotherapy.

[0281] Examples of anti-immune agents include, but are not limited to, adrenocortical hormones (e.g., fluticasone propionate, beclomethasone dipropionate, mometasone furoate, triamcinolone acetonide, or budesonide), disease-modifying agents (e.g., antimalarials, methotrexate, sulfasalazine salicylate, masalazone, azathioprine, 6-mercaptopurine, metronidazole, D-penicillamine), nonsteroidal anti-inflammatory drugs (e.g., acetaminophen, aspirin, sodium salicylate, sodium chlorambucil, magnesium salicylate, choline, magnesium salicylate, Salicylate, ibuprofen, naproxen, diclofenac, diflunisal, etodolac, fenoprofen calcium, flurbiprofen, piroxicam, indomethacin, ketoprofen, ketorolac tromethamine, meclofenamic acid base combination, meclofenamic acid sodium, mefenamic acid, naproxen, oxaprozin, butylphenyl nitrone, sulindac, or toluidine pyridine acetate), COX-2 inhibitors, cytokine synthesis / release inhibitors (such as anti-cytokine antibodies, anti-cytokine receptor antibodies, etc.).

[0282] Therefore, the methods described herein are not limited to the order of administration, and one or more other active ingredients may be administered simultaneously or before or after administration. In the combination described above, at least one pharmaceutically active ingredient is derived from a salt of formula A or a solvate thereof or a crystalline form of a salt of formula A or a solvate thereof (e.g., Form AI, Form BI, Form B-II, Form B-III, Form CI, Form C-II, and Form C-III).

[0283] The following are non-limiting examples.

[0284] Experimental Section

[0285] The compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine used in the examples was prepared according to WO2012167733A1.

[0286] All reagents (except intermediates) used in the present invention are purchased from the commercial market. All compound names (except reagents) are generated by software ChemBioDraw Ultra 16.0.

[0287] Unless otherwise noted, powder X-ray diffraction spectra were measured by Bruker D8 ADVANCE (target: Cu, voltage: 40 kV, current: 40 mA, scanning speed: 4 degrees / min, step size: 0.02 degrees, measurement range: 3-45 degrees).

[0288] Unless otherwise noted, differential scanning calorimetry was performed using a DSC 204F1 from NETZSCH (Germany) (purge gas: nitrogen, flow rate: 20-60 mL min -1 , heating rate: 5-10℃ / min, measuring range: 30℃→300℃), a rolled aluminum pan was used for sample measurement, and indium was used for temperature correction.

[0289] Unless otherwise noted, thermogravimetric analysis was performed using TGA 209F1 (purging gas: nitrogen, heating rate: 10°C / min) from NETZSCH, Germany.

[0290] Figure 8 and Fig.10 The differential scanning calorimetry and thermogravimetric analysis were measured by DSC Q2000 and TG Q500, respectively, from TA Company, USA.

[0291] Example 1: Preparation of Form AI of Formula A Salt

[0292] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine (100 mg, 0.21 mmol) was suspended in 1 ml of ethanol, heated to slight reflux under stirring, and 0.315 ml of 1M acetic acid aqueous solution was added thereto. The solution became clear and heating was stopped. The mixture was stirred at room temperature for 5 days, filtered, and dried to obtain 71.52 mg of a yellow solid.

[0293] 1 H NMR(400MHz,cdcl3)δ8.88(d,J=1.9Hz,1H),8.59(d,J=1.9Hz,1H),8.12(d,J=8.3Hz,2H),7.53(s,1H), 7.33(d,J=8.3Hz,2H),7.07(t,J=5.6Hz,1H),4.06–3.94(m,4H),3.89(tdd,J=6.6,3.9,2.4Hz,1H),3.70 (dtd,J=10.9,6.9,4.5Hz,2H),3.11(dd,J=12.3,2.0Hz,2H),2.93(dd,J=10.4,3.0Hz,2H),2.85(d,J=1. 7Hz,3H),2.83–2.75(m,2H),2.74–2.63(m,1H),2.05(s,2H),2.02(d,J=13.9Hz,2H),1.97–1.83(m,2H).

[0294] The obtained powder sample is the crystal form AI of the salt of formula A, and its powder X-ray diffraction pattern is as follows: Figure 1 The peaks selected have the following values: 6.2, 9.4, 10.8, 12.1, 14.1, 15.8, 16.3, 17.2, 19.0, 19.3, 20.1, 21.1, 21.7, 22.2, 22.7, 24.6, 25.4, 26.8, 27.2, 27.5, 29.2, 30.6 and 31.8 degrees, each with a different angle error of ±0.2 degrees (2θ), with characteristic peaks at 6.2, 9.4, 16.3, 17.2 and 19.0 degrees. The DSC test results are shown in Figure 2 As shown, Form AI has obvious endothermic peaks at about 162.8-179.6°C and about 217.0-219.4°C.

[0295] Example 2: Preparation of Form AI of Formula A Salt

[0296] Add glacial acetic acid (0.124 ml, 2.1 mmol) and (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine (100 mg, 0.21 mmol) to 2 ml of ethanol, heat and dissolve under stirring, stop heating, add 0.5 ml of isopropyl ether, continue stirring at room temperature overnight, filter, and dry to obtain 60 mg of yellow solid. After testing, the X-ray powder diffraction spectrum of the obtained sample is consistent with the crystal form AI sample of the salt of formula A obtained in Example 1.

[0297] Example 3: Preparation of Form AI of Formula A Salt

[0298] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine (100 mg, 0.21 mmol) was mixed with 0.5 ml of 1M acetic acid aqueous solution in 95% ethanol (5 ml), heated and dissolved under stirring, and the temperature of the mixture was lowered to room temperature, and 0.2 ml of isopropyl ether was added thereto, stirred at room temperature overnight, filtered, and dried to obtain 20 mg of a yellow solid. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form AI sample of the salt of formula A obtained in Example 1.

[0299] Example 4: Preparation of Form AI of Formula A Salt

[0300] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine (1.0 g, 2.1 mmol) was added to 4 ml of ethanol, heated to 50-55°C under stirring, and 3 ml of glacial acetic acid was added thereto to obtain a clear solution. 9 ml of methyl tert-butyl ether was added thereto dropwise, kept at 50-55°C for 30 minutes, then cooled to 20-25°C, stirred for 17 hours, filtered, and vacuum dried at 55°C for 5 hours to obtain 0.98 g of the product, with a yield of 87% and a purity of 99.55%. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form AI sample of the salt of formula A obtained in Example 1.

[0301] Example 5: Preparation of Form AI of Formula A Salt

[0302] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine (4.0 g, 8.29 mmol) was added to a mixed solvent formed by 16 ml of ethanol and 10 ml of water, and heated to 70-75°C under stirring. 1.42 ml of glacial acetic acid was added thereto to obtain a clear solution. 100 ml of ethanol was slowly added thereto, and the mixture was kept at 70-75°C for 30 minutes under stirring, and then slowly cooled to 0-5°C. The precipitated solid was filtered and vacuum dried at 55°C for 17 hours to obtain 3.8 g of the product, with a yield of 84% and a purity of 99.77%. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form AI sample of the salt of formula A obtained in Example 1.

[0303] Example 6: Preparation of Form AI of Formula A Salt

[0304] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine (1.0 g, 2.1 mmol) was added to 4 ml of isopropanol, heated to 50-55°C under stirring, and 3 ml of glacial acetic acid was added thereto to obtain a clear solution. 9 ml of methyl tert-butyl ether was added dropwise thereto, kept at 50-55°C for 30 minutes, then cooled to 20-25°C, stirred for 17 hours, filtered, and vacuum dried at 55°C for 5 hours to obtain 0.99 g of the product, with a yield of 88% and a purity of 99.54%. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form AI sample of the salt of formula A obtained in Example 1.

[0305] Example 7: Preparation of Form AI of Formula A Salt

[0306] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine (48 mg, 0.1 mmol) was suspended in 4 ml of isopropanol, heated to 80°C with stirring, and 0.32 ml of 1M acetic acid aqueous solution was added thereto to obtain a clear solution. After cooling to room temperature, the mixture was stirred overnight, filtered, and dried to obtain 40 mg of a solid. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form AI sample of the salt of formula A obtained in Example 1.

[0307] Example 8: Preparation of Form AI of Formula A Salt

[0308] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine (1.0 g, 2.1 mmol) was suspended in 10 ml of isopropanol, heated to 80-85°C, and 3.11 ml of 1M aqueous acetic acid solution was added to obtain a clear solution. 10 ml of isopropanol was added thereto, and the temperature of the reaction system was lowered to room temperature and stirred overnight. The solid was collected by filtration, the filter cake was washed with isopropanol, and vacuum dried at 60°C for 2 hours to obtain 880 mg of the product with a purity of 99.2%. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form AI sample of the salt of formula A obtained in Example 1.

[0309] Example 9: Preparation of Form AI of Formula A Salt

[0310] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine (100 mg, 0.21 mmol) was suspended in 1 ml of acetonitrile, heated to slight reflux under stirring, 0.315 ml of 1M acetic acid aqueous solution was added thereto, the solution became clear, heating was stopped, stirring was continued at room temperature overnight, 1 ml of acetonitrile was added thereto, stirring was continued for 5 days, filtering, and drying to obtain 38.54 mg of a yellow solid. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form AI sample of the salt of formula A obtained in Example 1.

[0311] Example 10: Preparation of Form AI of Formula A Salt

[0312] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine (48 mg, 0.1 mmol) was suspended in 4.5 ml of acetonitrile, heated to 80-85°C, 0.2 ml of 1M acetic acid aqueous solution and 0.1 ml of water were added to obtain a clear solution, and then the temperature of the reaction system was lowered to room temperature and stirred overnight. The solid was collected by filtration and dried to obtain 39.4 mg of the product. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form AI sample of the salt of formula A obtained in Example 1.

[0313] Example 11: Preparation of Form AI of Formula A Salt

[0314] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine (100 mg, 0.21 mmol) was suspended in 1 ml of acetone, heated to slight reflux, 0.315 ml of 1M acetic acid aqueous solution was added to obtain a clear solution, and heating was stopped. The reaction system was stirred at room temperature overnight, the product precipitated, and stirring at room temperature was continued for 5 days. The solid was collected by filtration and dried at room temperature to obtain 76.04 mg of the product. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form AI sample of the salt of formula A obtained in Example 1.

[0315] Example 12: Preparation of Form AI of Formula A Salt

[0316] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine (1.0 g, 2.1 mmol) was dissolved in 3 ml of glacial acetic acid at 60-64°C, 18 ml of tert-butyl alcohol was added dropwise, and stirring was continued for 30 minutes; after cooling to 20-25°C, stirring was continued for 17 hours, and the solid was collected by filtration and vacuum dried at 60°C for 6 hours to obtain 1.04 g of the product, with a yield of 92% and a purity of 98.74%. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form AI sample of the salt of formula A obtained in Example 1.

[0317] Example 13: Preparation of Form AI of Formula A Salt

[0318] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine (1.0 g, 2.1 mmol) was suspended in a mixed solvent consisting of 6 ml of dioxane and 2 ml of water, heated to 45-50°C with stirring, and 0.36 ml of glacial acetic acid was added to obtain a clear solution. 10 ml of dioxane was added thereto at 50-55°C. The reaction system was cooled to 20-25°C and continued to stir for 2 hours. The solid was collected by filtration and dried in vacuo at 55°C to obtain 0.7 g of the product, with a yield of 62% and a purity of 99.83%. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form AI sample of the salt of formula A obtained in Example 1.

[0319] Example 14: Preparation of Form AI of Formula A Salt

[0320] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine (1.0 g, 2.1 mmol) was dissolved in 3 ml of glacial acetic acid at 40-50°C, 35 ml of dioxane was added dropwise thereto, after addition, the reaction system was cooled to 20-25°C, stirred for 1 hour, and then cooled to 5-10°C and stirred for 1 hour, the solid was collected by filtration, and vacuum dried at 55°C for 17 hours to obtain 0.23 g of the product. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form AI sample of the salt of formula A obtained in Example 1.

[0321] Example 15: Preparation of Form AI of Formula A Salt

[0322] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine (0.5 g, 1.04 mmol) was dissolved in 9.5 ml of dichloromethane at 40-45°C, 0.12 ml of glacial acetic acid was added thereto under stirring, and then 3 ml of methyl tert-butyl ether was added, and the mixture was stirred at 40-45°C for 10 minutes, then cooled to 20-25°C and stirred for 17 hours. The solid was collected by filtration and dried under vacuum at 55°C for 4 hours to obtain 0.5 g of the product with a purity of 99.56%. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form AI sample of the salt of formula A obtained in Example 1.

[0323] Example 16: Preparation of Form AI of Formula A Salt

[0324] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine (48 mg, 0.1 mmol) was suspended in 1 ml of methanol, heated to 70°C, and 0.2 ml of 1M acetic acid aqueous solution was added to obtain a clear solution. The temperature of the reaction system was lowered to room temperature and stirred overnight. The solid was collected by filtration and dried to obtain 30.2 mg of the product. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form AI sample of the salt of formula A obtained in Example 1.

[0325] Example 17: Preparation of Form AI of Formula A Salt

[0326] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine (1.5 g, 3.11 mmol) and 0.37 g of glacial acetic acid were suspended in a mixed solvent consisting of 6 ml of acetone and 3 ml of water, heated to 50°C, stirred at this temperature for 20 minutes, and then 27 ml of acetone was slowly added thereto, the temperature was raised to 60°C and stirred for 2 hours, then naturally cooled to room temperature and stirred overnight, the solid was collected by filtration, and dried to obtain 1.4 g of the product with a purity of 99.72%. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form AI sample of the salt of formula A obtained in Example 1.

[0327] Example 18: Preparation of Form BI of Formula A Salt

[0328] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine (1.0 g, 2.1 mmol) was suspended in a mixed solvent consisting of 30 ml of isopropanol and 4.6 ml of water, heated to 80-85°C with stirring, and 2.3 ml of 1M p-toluenesulfonic acid aqueous solution was added. After the reaction system was dissolved, the temperature was lowered to 15-20°C and stirring was continued for 16 hours. The solid was collected by filtration and dried to obtain 1.06 g of the product.

[0329] 1 H NMR(400MHz,dmso)δ8.03–7.99(m,J=2.0Hz,1H),7.76–7.73(m,J=2.0Hz,1H),7.18–7.11(m,J=8.4Hz,2H),7.06–6.99(m,J=6.0 Hz,1H),6.58–6.55(m,1H),6.47–6.42(m,2H),6.41–6.36(m,J=8.4Hz,2H),6.10–6.05(m,J=7.8Hz,2H),3.13–3.04(m,1H),3.0 4–2.96(m,J=12.5,3.2Hz,1H),2.83–2.64(m,7H),2.18–2.11(m,J=12.2Hz,1H),2.08–1.96(m,J=12.5,8.7Hz,1H),1.89(s,3H) ,1.86–1.78(m,J=12.0,9.6Hz,3H),1.74–1.66(m,J=12.0Hz,1H),1.25(s,3H),0.94–0.85(m,J=12.6Hz,2H),0.77–0.64(m,2H).

[0330] The obtained powder sample is the crystal form BI of the salt of formula A, and its powder X-ray diffraction pattern is as follows: Figure 4 The peaks selected have the following values: 4.9, 5.5, 6.8, 9.6, 10.1, 14.4, 14.8, 15.9, 16.4, 17.3, 18.4, 19.3, 19.8, 20.7, 21.4, 22.5, 23.2, 25.0, 26.1, 27.0 and 29.0 degrees, each with an angle error of ±0.2 degrees (2θ), of which the characteristic peaks are 4.9, 5.5, 9.6, 14.4, 16.4 and 19.8 degrees. The DSC test results are shown in Figure 5 As shown, the endothermic peak of Form BI is at about 205.7-209.6°C.

[0331] Example 19: Preparation of Form BI of Formula A Salt

[0332] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine (48 mg, 0.1 mmol) was suspended in 1.5 ml of 95% ethanol at 80°C, and 0.11 ml of 1M p-toluenesulfonic acid aqueous solution was added thereto to obtain a clear solution. The reaction system was cooled to 15-20°C and stirred for 3 hours. The product was filtered and dried. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystalline form BI sample of the salt of formula A obtained in Example 18.

[0333] Example 20: Preparation of Form BI of Formula A Salt

[0334] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine (100 mg, 0.21 mmol) was suspended in 1 ml of ethanol, heated to slight reflux under stirring, 0.23 ml of 1M p-toluenesulfonic acid aqueous solution was added thereto, the solution became clear, heating was stopped to precipitate oil, 2 ml of ethanol was added and stirring was continued at room temperature for 3 days, filtered, and dried to obtain 124.36 mg of solid. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form BI sample of the salt of formula A obtained in Example 18.

[0335] Example 21: Preparation of Form BI of Formula A Salt

[0336] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine (100 mg, 0.21 mmol) was suspended in a mixed solvent consisting of 2 ml of ethanol and 0.23 ml of water, heated to 80-90°C, and 0.23 ml of 1M p-toluenesulfonic acid aqueous solution was added thereto to obtain a clear solution. Cool to room temperature and continue stirring for 16 hours. The solid was collected by filtration and dried at room temperature to obtain 85 mg of the product. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystalline form BI sample of the salt of formula A obtained in Example 18.

[0337] Example 22: Preparation of Form BI of Formula A Salt

[0338] About 50 mg of the crystal form B-II sample of the salt of formula A was suspended in about 0.5 ml of 95% ethanol and stirred at 20° C. for 4 days, and the solid was collected by filtration to obtain the product. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form BI sample of the salt of formula A obtained in Example 18.

[0339] Example 23: Preparation of Form BI of Formula A Salt

[0340] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine (100 mg, 0.21 mmol) was suspended in 1 ml of isopropanol, heated to slight reflux under stirring, 0.23 ml of 1M p-toluenesulfonic acid aqueous solution was added thereto, the solution became clear, heating was stopped to precipitate an oily substance, 3 ml of isopropanol was added and stirring was continued at room temperature for 3 days, filtered, and dried to obtain 131.92 mg of solid. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form BI sample of the salt of formula A obtained in Example 18.

[0341] Example 24: Preparation of Form BI of Formula A Salt

[0342] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine (100 mg, 0.21 mmol) was suspended in 1 ml of acetone, heated to slight reflux under stirring, 0.23 ml of 1M p-toluenesulfonic acid aqueous solution was added thereto, the solution became clear, heating was stopped to precipitate an oily substance, 4 ml of acetone was added and stirring was continued at room temperature for 3 days, filtered, and dried to obtain 94.77 mg of solid. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form BI sample of the salt of formula A obtained in Example 18.

[0343] Example 25: Preparation of Form B-II of the Salt of Formula A

[0344] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine (100 mg, 0.21 mmol) was suspended in 1 ml of methanol and heated to slight reflux with stirring. 0.23 ml of 1M aqueous p-toluenesulfonic acid solution was added thereto. The solution became clear. Heating was stopped to precipitate an oily substance. After adding 2 ml of methanol, stirring was continued at room temperature for 3 days. The mixture was filtered and dried to obtain 71.66 mg of a solid.

[0345] 1 H NMR (400MHz, cdcl3) δ8.88(d,J=1.9Hz,1H),8.57(d,J=1.9Hz,1H),8.07(d,J=8.4Hz,2H),7.65(d,J=8. 2Hz,2H),7.55(s,1H),7.31(d,J=8.3Hz,2H),7.07(d,J=7.9Hz,2H),6.95(t,J=6.0Hz,1H),4.21–4.12(m ,1H),4.04–3.88(m,6H),3.47(dd,J=42.8,12.1Hz,2H),3.12–3.03(m,1H),2.98(t,J=11.8Hz,1H),2.8 3(s,3H),2.80–2.75(m,2H),2.71–2.62(m,1H),2.28(s,3H),1.98(d,J=12.4Hz,2H),1.92–1.84(m,2H).

[0346] The obtained powder sample is the crystalline form B-II of the salt of formula A, and its powder X-ray diffraction pattern is as follows: Figure 7 The peaks selected have the following values: 5.1, 6.0, 9.5, 10.2, 14.3, 14.8, 15.3, 15.8, 17.1, 17.9, 19.1, 19.7, 20.2, 20.8, 22.4, 23.4, 26.0 and 27.2 degrees, each with an angle error of ±0.2 degrees (2θ), with characteristic peaks at 5.1, 6.0, 10.2, 17.1 and 19.1 degrees. The DSC test results are shown in Figure 8 As shown, the endothermic peak of Form B-II is at about 203.0-211.4°C.

[0347] Example 26: Preparation of Form B-III of the Salt of Formula A

[0348] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine (48 mg, 0.1 mmol) was suspended in 2.5 ml of water at 80-85°C, and 0.11 ml of 1 M p-toluenesulfonic acid aqueous solution was added thereto to obtain a clear solution. The solution was cooled to room temperature with stirring to precipitate, which was filtered and dried to obtain a solid.

[0349] The obtained powder sample is the crystalline form B-III of the salt of formula A, and its powder X-ray diffraction pattern is as follows: Fig. 9 The peaks selected have the following values: 5.3, 5.9, 9.9, 10.7, 11.8, 13.6, 14.9, 15.6, 16.0, 17.6, 18.9, 20.0, 21.6, 22.8, 25.0 and 27.0 degrees, each with a different angle error of ±0.2 degrees (2θ), of which the characteristic peaks are 5.3, 5.9, 10.7, 13.6 and 15.6 degrees. The DSC test results are shown in Fig.10 As shown, Form B-III has an exothermic peak at about 168.1-172.5°C and an obvious endothermic peak at about 199.6-208.3°C.

[0350] Example 27: Preparation of Form B-III of the Salt of Formula A

[0351] About 50 mg of the crystal form BI sample of the salt of formula A was suspended in 0.5 ml of water, stirred at room temperature for 4 days, filtered to collect the solid, and vacuum dried at 60° C. for 2 hours to obtain the product. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form B-III sample of the salt of formula A obtained in Example 26.

[0352] Example 28: Preparation of Form CI of Formula A Salt

[0353] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine (96.5 mg, 0.2 mmol) was suspended in 3 ml of isopropanol and heated to 80-85°C with stirring. 0.3 ml of 1M malic acid aqueous solution and 1.8 ml of water were added thereto, and the reaction system became a clear solution. Cooled to room temperature and stirred overnight. The solid was collected by filtration and dried to obtain 97.7 mg of the product.

[0354] 1H NMR(400MHz,dmso)δ9.03(d,J=2.0Hz,1H),8.77(d,J=2.0Hz,1H),8.17(d,J=8. 4Hz,2H),7.98(t,J=6.1Hz,1H),7.58(s,1H),7.41(d,J=8.4Hz,2H),4.07–3.94( m,3H),3.91(dd,J=9.8,4.2Hz,1H),3.78–3.61(m,8H),3.07–2.66(m,14H),2.3 3(dd,J=15.6,4.2Hz,1H), 1.93(d,J=13.2Hz,2H), 1.74(dt,J=12.5,8.9Hz,2H).

[0355] The obtained powder sample is the crystal form CI of the salt of formula A, and its powder X-ray diffraction pattern is as follows: Fig.11 The peaks selected have the following values: 8.6, 10.8, 11.8, 13.5, 14.3, 15.5, 16.4, 17.2, 17.7, 18.1, 18.4, 19.5, 20.9, 22.2, 22.6, 23.8, 25.8, 26.7, 27.8, 29.2, 29.8 and 30.7 degrees, each with an angle error of ±0.2 degrees (2θ), with characteristic peaks at 8.6, 14.3, 15.5, 19.5 and 22.2 degrees. The DSC test results are shown in Fig.12 As shown, the endothermic peak of the crystal form CI is about 211.1-214.9°C. Example 29: Preparation of the crystal form CI of the salt of formula A

[0356] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazine-5-amine (1.0 g, 2.1 mmol) was suspended in 20 ml of ethanol, heated to 80°C, 3.1 ml of 1M malic acid aqueous solution was added thereto, and then 12 ml of water was added, and the reaction system became a clear solution. Then 3 ml of ethanol was added, the temperature of the reaction system was cooled to 15-20°C, and stirring was continued for 14 hours. Filtered, vacuum dried at 60°C for 2 hours, and 1.03 g of the product was obtained with a purity of 99.7%. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form CI sample of the salt of formula A obtained in Example 28.

[0357] Example 30: Preparation of Form CI of Formula A Salt

[0358] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine (96.5 mg, 0.2 mmol) was mixed with 7 ml of tetrahydrofuran, 0.7 ml of water and 0.3 ml of 1M malic acid aqueous solution and heated to dissolve to obtain a clear solution, and then the reaction system was cooled to 15-20°C, 5 ml of tetrahydrofuran was added, and stirring was continued for 3 days, filtered, and dried to obtain 87.9 mg of the product. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form CI sample of the salt of formula A obtained in Example 28.

[0359] Example 31: Preparation of Form CI of Formula A Salt

[0360] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine (96.5 mg, 0.2 mmol) was mixed with 5 ml of methanol, 3 ml of water and 0.3 ml of 1M malic acid aqueous solution and heated to 75-80°C to obtain a clear solution, and then the reaction system was slowly cooled to 15-20°C under stirring, filtered, and dried to obtain 104 mg of the product. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form CI sample of the salt of formula A obtained in Example 28.

[0361] Example 32: Preparation of Form CI of Formula A Salt

[0362] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine (48 mg, 0.1 mmol) was suspended in 3 ml of ethanol, and 14.8 mg of malic acid suspended in 0.2 ml of ethanol was added thereto. The reaction system was stirred at 40°C for 20 hours, cooled to room temperature under stirring, filtered, and dried to obtain about 40 mg of the product. After testing, the X-ray powder diffraction spectrum of the obtained sample was consistent with the crystal form CI sample of the salt of formula A obtained in Example 28.

[0363] Example 33: Preparation of Form C-II of the Salt of Formula A

[0364] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine (48 mg, 0.1 mmol), 3.5 ml of water, 0.6 ml of acetonitrile and 0.11 ml of 1M malic acid aqueous solution were mixed and heated to 80-85°C under stirring to dissolve, and then the reaction solution was cooled to room temperature and stirred for 18 hours. Filtered and dried to obtain 35.2 mg of the product.

[0365] 1H NMR(400MHz,dmso)δ9.02(d,J=2.0Hz,1H),8.77(d,J=2.0Hz,1H),8.17(d,J=8.4Hz,2H),7.89(t,J=6.0Hz,1H),7.57 (s,1H),7.42(d,J=8.4Hz,2H),3.89(d,J=9.1Hz,2H),3.84(dd,J=10.8,3.4Hz,1H),3.80–3.63(m,5H),3.56(dd,J=2 3.4,11.8Hz,2H),3.08(d,J=12.7Hz,1H),2.92(d,J=2.1Hz,4H),2.85(dd,J=12.2,9.8Hz,5H),2.70(dt,J=22.5,11. 2Hz, 3H), 2.46 (d, J = 10.8Hz, 1H), 2.31 (dd, J = 15.6, 3.4Hz, 1H), 1.93 (d, J = 12.9Hz, 2H), 1.74 (dt, J = 12.2, 8.5Hz, 2H).

[0366] The obtained powder sample is the crystal form C-II of the salt of formula A, and its powder X-ray diffraction pattern is as follows: Fig.14 The peaks selected have the following values: 5.3, 9.5, 10.0, 10.6, 11.3, 11.6, 12.4, 12.8, 13.6, 15.9, 17.0, 18.4, 20.0, 20.9, 21.6, 23.1, 24.9, 26.4, 26.9 and 28.3 degrees, each with an angle error of ±0.2 degrees (2θ), and the characteristic peaks are 11.3, 11.6, 17.0, 20.0, 21.6 and 23.1 degrees. The DSC test results are shown in Fig.15 As shown, the endothermic peaks of Form C-II are at about 70.2-90.4°C and about 202.6-215.0°C.

[0367] Example 34: Preparation of Form C-III of the Salt of Formula A

[0368] (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine (1.0 g, 2.1 mmol), 11 ml of ethanol, 41 ml of water and 3.1 ml of 1M malic acid aqueous solution were mixed and heated to 80°C to dissolve, and then the reaction system was slowly cooled to 15-20°C and continued to stir overnight. The solid was collected by filtration and dried to obtain 0.94 g of the product with a purity of 99.7%.

[0369] 1H NMR(400MHz,dmso)δ9.02(d,J=2.0Hz,1H),8.76(d,J=2.0Hz,1H),8.16(d,J=8.4Hz,2H),7.90(t,J=5. 9Hz,1H),7.57(s,1H),7.41(d,J=8.4Hz,2H),3.90(d,J=8.9Hz,2H),3.84(dd,J=10.7,3.4Hz,1H),3.8 0–3.65(m,5H),3.62–3.54(m,2H),3.10(d,J=11.8Hz,2H),2.98–2.79(m,7H),2.78–2.65(m,2H),2.46 (d, J=10.7Hz, 1H), 2.31 (dd, J=15.6, 3.4Hz, 1H), 1.92 (d, J=12.8Hz, 2H), 1.74 (qd, J=12.6, 4.0Hz, 2H).

[0370] The obtained powder sample is the crystal form C-III of the salt of formula A, and its powder X-ray diffraction pattern is as follows: Fig.17 The peaks selected have the following values: 5.3, 10.6, 11.8, 12.8, 16.0, 17.0, 17.9, 18.6, 18.9, 20.3, 21.7, 22.5, 23.1, 25.2, 25.6 and 27.4 degrees, each with an angle error of ±0.2 degrees (2θ), of which the characteristic peaks are 5.3, 10.6, 17.0, 17.9 and 25.2 degrees. The DSC test results are shown in Fig.18 As shown, the endothermic peaks of Form C-III are at about 69.9-88.5°C and about 201.6-206.7°C.

[0371] Example 35: Stability test of salt of formula A

[0372] Determination method: Take the test samples of the crystal form AI, crystal form BI, crystal form CI and crystal form C-III of the salt of formula A respectively and place them in a culture dish, and place them in a sealed clean container with the opening exposed. Place them at a temperature of 60°C for 10 days, at an illumination of 4500lx±500lx for 10 days, and at 25°C and a relative humidity of 92.5%±5% for 10 days. Take samples on the 5th day and the 10th day respectively, and examine the purity (by HPLC analysis) and crystal form (by X-ray powder diffraction analysis) of the samples, and compare the inspection results. The experimental results of various salts and their crystal forms are shown in Tables 1, 2, 3 and 4.

[0373] Table 1: Stability test results of the salt of formula A, form AI

[0374]

[0375] Table 2: Stability test results of the salt of formula A, form BI

[0376]

[0377] Table 3: Stability test results of the salt of Formula A, Form CI

[0378]

[0379]

[0380] Table 4: Stability test results of the salt of formula A, form C-III

[0381]

[0382] Conclusion: The data in Tables 1, 2, 3 and 4 show that the chemical purity and crystal form of the salt of Formula A, Form AI, Form BI, Form CI and Form C-III, are not changed when placed under high temperature conditions for 10 days, under light conditions for 10 days or under high humidity conditions for 10 days, indicating that the salt of Formula A, Form AI, Form BI, Form CI and Form C-III are stable.

[0383] Example 36: Solubility test of salt of formula A

[0384] (1) Determination of the solubility of the salt of Formula A (acetate, crystal form AI): Appropriate amounts of the salt of Formula A (acetate, crystal form AI) and its free base (compound (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine) were added to various solvents, stirred at a certain temperature for 4 hours, the excess solid was filtered off, and the clear filtrate was taken to determine the solubility. The experimental results are shown in Table 5.

[0385] Table 5

[0386]

[0387]

[0388] Conclusion: The data in Table 5 show that the solubility of the salt of Formula A (acetate, Form AI) in water is significantly increased compared with its free base.

[0389] (2) Rough solubility determination of the salt of formula A: The salt of formula A to be roughly solubilized was weighed in advance and added to a test tube, and then pure water was added dropwise to the test tube in batches, and the amount of water added each time was recorded. After each addition of water, the test tube was shaken for 5 minutes until all the solid was dissolved. The rough solubility of the compound was then calculated based on the mass of the weighed compound and the total amount of water added. The calculation results are shown in Table 6.

[0390] Table 6

[0391] Dissolution medium Test temperature Formula A salt Rough solubility (mg / mL) Pure water 19℃ Acetate (Crystal Form AI) 14.6 Pure water 19℃ p-Toluenesulfonate (Form BI) 0.65 Pure water 19℃ Malate (Form CI) 0.48

[0392] Conclusion: The data in Table 6 combined with the data in Table 5 show that the solubility of the salt of Formula A in water is increased to varying degrees compared with its free base, among which the solubility of the acetate salt is particularly increased, followed by the p-toluenesulfonate and malate salts.

[0393] It should be understood that the embodiments and implementations described herein are for illustrative purposes only, and various modifications or changes thereto will be suggested to those skilled in the art, which are included within the spirit and scope of the present application and the scope of the appended claims. All publications, patents and patent applications cited herein are incorporated herein by reference and used for all purposes.

Claims

1. Crystalline form BI of p-toluenesulfonate of (S)-7-(4-(1-(methylsulfonyl)-piperidin-4-yl)phenyl)-N-(morpholin-2-ylmethyl)pyrido[3,4-b]pyrazin-5-amine, characterized in that: The crystalline form BI has characteristic peaks at the following 2θ angles in the powder X-ray diffraction pattern: 4.9 degrees, 5.5 degrees, 9.6 degrees, 14.4 degrees, 16.4 degrees and 19.8 degrees, and each 2θ value has an error of about ±0.2°.

2. A pharmaceutical composition, characterized in that The pharmaceutical composition contains an effective amount of the crystalline form BI according to claim 1 and a pharmaceutically acceptable carrier.

3. Use of the crystalline form BI according to claim 1 in the preparation of a drug for treating a disease associated with Syk kinase activity.

4. The use according to claim 3, wherein the disease is an autoimmune disease, an inflammatory disease or cancer.

5. The use according to claim 3, wherein the disease is a hematological malignancy.

6. The method of claim 4, wherein the autoimmune disease, inflammatory disease and cancer are systemic lupus erythematosus, rheumatoid arthritis, Sjögren's syndrome, IgA nephropathy, autoimmune hemolytic anemia, idiopathic thrombocytopenic purpura, multiple sclerosis, myasthenia gravis, allergic rhinitis, chronic obstructive pulmonary disease, adult respiratory distress syndrome, asthma, lymphoma, leukemia or multiple myeloma.

7. The use according to claim 6, wherein the lymphoma is a B-cell lymphoma or a T-cell lymphoma.

8. The use according to claim 6, wherein the leukemia is chronic lymphocytic leukemia, acute lymphocytic leukemia or acute myeloid leukemia.

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